Nitrogen pollution is a growing problem that is detrimental to the environment and the economy. Traditional treatment of nitrogen is a multi-stage process, expensive, operationally intensive, and requires large land areas. This research studied the effects of BiOWiSH® Aqua (Aqua), a biological enhancement product, on the simultaneous nitrification and denitrification process in a membrane aerated bioreactor (MABR) to determine if it is a feasible application for wastewater treatment. The MABR used during experimentation was a small-scale batch reactor with a continuous flow of air through a silicone membrane. The effect of carbon source and concentration on nitrogen removal rates and biomass growth/behavior were determined through a series of laboratory experiments with Aqua and wastewater. With glucose and solely Aqua cultures, average reduction rates in nitrogen concentrations were 1.2 mg-N/L/hour for all C:N ratios investigated. When wastewater was used as the main carbon source, creating a mix of wastewater and Aqua bacteria in the MABR, average reduction rates were 10.9 mg-N/L/hour. A maximum reduction rate of 21.3 mg-N/L/hour occurred at a 2:1 C:N ratio. This research concluded that pure Aqua cultures are not efficient at removing nitrogen or greatly augment the nitrogen reduction process. MABRs can use the biochemical oxygen demand in wastewater as a useful/viable carbon source. High carbon to nitrogen ratios (C:N ratio of 30:1) did not result in faster nitrogen reduction rates but did experience rapid biofilm growth and death. This shows that high C:N ratios are not an efficient operationally for MABRs due to the excess sludge created. C:N ratios of v approximately 3:1 provided the most consistent nitrogen reduction for both glucose and wastewater. This research concluded that C:N ratios, pH, and oxygen diffusion heavily affect the MABR’s performance. In addition, MABRs can utilize low C:N ratios during treatment, particularly during the treatment of high-strength wastewater.

The membrane aerated bioreactor (MABR) is a unique technological innovation where a gas permeable membrane is applied to biological processes. In an MABR, oxygen and other substrates diffuse from the opposite directions into a biofilm, and thus simultaneous chemical oxygen demand (COD) and nitrogen removal can be achieved. However, controlling biofilm thickness, stability, and attachment is challenging. The objectives of this research were to study the effect of oxygen partial pressure on process performance with respect to nitrogen removal and examine the biomass properties in MABRs at different oxygen partial pressures and COD loadings. The conditions within the bioreactors were based on a low hydrodynamic condition (average fluid velocity 22 cm/min along the membrane surface), with the intention of minimizing the impact of the hydrodynamic shear on biomass properties. Simultaneous nitrification and denitrification were achieved in the reactors, and increasing oxygen partial pressure enhanced the total nitrogen removal. The biomass at the membrane-biofilm interface was more porous at a loading of 11.3 kg COD/1000 m2/day (areal porosity about 0.9) as compared with a loading of 22.6 kg COD/1000 m2/day (areal porosity about 0.7), indicating carbon substrate was limiting near the membrane. Long-term (over 30 days) experimental results showed that at the loading of 11.3 kg COD/1000 m2/day, the oxygen partial pressures of 0.59 atm and 0.88 atm caused over 80% of the biomass to become suspended in the bulk phase while at 0.25 atm and 0.41 atm oxygen over 97% of the biomass was immobilized on the membrane. There is a critical oxygen partial pressure that can sustain the biofilm, which increases with an increasing COD loading. The nitrifying population in the reactors was examined by applying fluorescence in situ hybridization (FISH). At the loading of 22.6 kg COD/1000 m2/day, there were 12% beta-proteobacterial ammonia oxidizing bacteria (AOB) and 17%Nitrobacter in homogenized biofilm biomass at 0.59 atm oxygen while there were 7% beta-proteobacterial AOB and 4% Nitrobacter at 0.25 atm oxygen. The ratio of protein to carbohydrate in extracellular polymeric substances (EPS) of the homogenized biomass in the reactor decreased with increasing oxygen partial pressure. Surface characterization of the biomass revealed that the higher the oxygen partial pressure, the lower the biomass hydrophobicity and surface charge. The ratio of EPS protein to carbohydrate in a membrane aerated biofilm decreased when approaching the membrane-biofilm interface. The distribution of nitrifiers and dissolved oxygen profiles inside the biofilm suggested that dual substrate limitations exist, and it was concluded that the membrane aerated biofilm had an aerobic region in the inner layer and an anoxic region in the outer layer. It is proposed that the loss of EPS due to secondary substrate consumption, especially the loss of EPS proteins, at the bottom of the biofilm was responsible for biofilm detachment subjected to a critical oxygen partial pressure.

The membrane aerated bioreactor (MABR) is a unique technological innovation where a gas permeable membrane is applied to biological processes. In an MABR, oxygen and other substrates diffuse from the opposite directions into a biofilm, and thus simultaneous chemical oxygen demand (COD) and nitrogen removal can be achieved. However, controlling biofilm thickness, stability, and attachment is challenging. The objectives of this research were to study the effect of oxygen partial pressure on process performance with respect to nitrogen removal and examine the biomass properties in MABRs at different oxygen partial pressures and COD loadings. The conditions within the bioreactors were based on a low hydrodynamic condition (average fluid velocity 22 cm/min along the membrane surface), with the intention of minimizing the impact of the hydrodynamic shear on biomass properties. Simultaneous nitrification and denitrification were achieved in the reactors, and increasing oxygen partial pressure enhanced the total nitrogen removal. The biomass at the membrane-biofilm interface was more porous at a loading of 11.3 kg COD/1000 m2/day (areal porosity about 0.9) as compared with a loading of 22.6 kg COD/1000 m2/day (areal porosity about 0.7), indicating carbon substrate was limiting near the membrane. Long-term (over 30 days) experimental results showed that at the loading of 11.3 kg COD/1000 m2/day, the oxygen partial pressures of 0.59 atm and 0.88 atm caused over 80% of the biomass to become suspended in the bulk phase while at 0.25 atm and 0.41 atm oxygen over 97% of the biomass was immobilized on the membrane. There is a critical oxygen partial pressure that can sustain the biofilm, which increases with an increasing COD loading. The nitrifying population in the reactors was examined by applying fluorescence in situ hybridization (FISH). At the loading of 22.6 kg COD/1000 m2/day, there were 12% beta-proteobacterial ammonia oxidizing bacteria (AOB) and 17%Nitrobacter in homogenized biofilm biomass at 0.59 atm oxygen while there were 7% beta-proteobacterial AOB and 4% Nitrobacter at 0.25 atm oxygen. The ratio of protein to carbohydrate in extracellular polymeric substances (EPS) of the homogenized biomass in the reactor decreased with increasing oxygen partial pressure. Surface characterization of the biomass revealed that the higher the oxygen partial pressure, the lower the biomass hydrophobicity and surface charge. The ratio of EPS protein to carbohydrate in a membrane aerated biofilm decreased when approaching the membrane-biofilm interface. The distribution of nitrifiers and dissolved oxygen profiles inside the biofilm suggested that dual substrate limitations exist, and it was concluded that the membrane aerated biofilm had an aerobic region in the inner layer and an anoxic region in the outer layer. It is proposed that the loss of EPS due to secondary substrate consumption, especially the loss of EPS proteins, at the bottom of the biofilm was responsible for biofilm detachment subjected to a critical oxygen partial pressure.

This research entails the investigation of the effects of a bioaugmentation product from BiOWiSH® called Aqua, referred to as “Aqua” for the remainder of this paper, on the nitrogen removal rate in a membrane aerated bioreactor (MABR). This research was conducted using a MABR design that consisted of a silicone membrane and continuous flow airline with compressed air. The membrane system was designed to supply oxygen, creating an aerated layer at the membrane-biofilm interface and an anoxic layer at the biofilm-water interface. Laboratory experiments were conducted to compare the nitrogen removal rates of natural bacteria alone to natural bacteria paired with Aqua. However, it was not possible to determine if a difference existed between the nitrogen removal rates of the MABR systems with only natural bacteria versus those with natural bacteria augmented with Aqua. The mean nitrogen removal rate observed when the media in the system reached steady state was 0.39 mg-N/L-hr. with a carbon to nitrogen (C: N) ratio of 12:1. The only increase in the nitrogen removal rate observed was when the C: N ratio was doubled to 24:1 and the nitrogen removal rate increased to 0.56 mg-N/L-hr. Although it appeared that the Aqua did not have an influence on the nitrogen removal rate in the MABR systems, many other variables still need to be assessed to reach a conclusion. To improve the efficiency of the system more tubing should be added, or the glucose should be removed from the growth media because the maximum O2 mass transfer rate is only enough O2 for nitrification. The addition of glucose at 12:1 ratio increased the O2 demand in the system to be five times greater than the O2 supplied from the silicone tubing. This research determined that use of trace minerals, Aqua dosing method, and Aqua dosing concentration were not contributing factors in nitrogen removal from growth media under the conditions of this experiment.