Patricia Reboleiro-Rivas

Bacterial community structure and enzyme activities in a membrane bioreactor (MBR) using pure oxygen as an aeration source

A pilot-scale membrane bioreactor was used to treat urban wastewater using pure oxygen instead of air as a source of aeration, to study its influence on bacterial diversity and levels of enzyme activities (acid and alkaline phosphatases, glucosidase, protease, and esterase) in the sludge. The experimental work was developed in two stages influenced by seasonal temperature. Operational parameters (temperature, pH, BOD5, COD, total and volatile suspended solids) were daily monitored, and enzyme activities measured twice a week. Redundancy analysis (RDA) was used to reveal relationships between the level of enzyme activities and the variation of operational parameters, demonstrating a significant effect of temperature and volatile suspended solids. Bacterial diversity was analyzed by temperature-gradient gel electrophoresis of PCR-amplified partial 16S rRNA genes. Significant differences in community structure were observed between both stages. Sequence analysis revealed that the prevalent Bacteria populations were evolutively close to Alphaproteobacteria (44%), Betaproteobacteria (25%) and Firmicutes (17%).

Kinetic study and oxygen transfer efficiency evaluation using respirometric methods in a submerged membrane bioreactor using pure oxygen to supply the aerobic conditions.

The performance of a wastewater bench-scale ultrafiltration membrane bioreactor (MBR) treatment plant using pure oxygen to supply the aerobic conditions for 95 days was studied. The results showed the capacity of the MBR systems to remove organic material under a hydraulic retention time of 12h and a sludge retention time of 39.91 days. Aeration represents its major power input; this is why the alpha-factor of the aeration and kinetic parameters (design parameters) were determined when the mixed liquid suspended solids (MLSS) was increased from 3420 to 12,600 mg/l in order to understand the system. An alpha-factor in the range 0.462-0.022 and the kinetic parameters measured with the respirometric method (K(M) of 73.954-3.647 mg/l, k(d) of 0.0142-0.104 day(-1), k(H) of 0.1266-0.655 day(-1), and the yield mean coefficient of 0.941) were obtained. Our study suggested significant changes in the behaviour of the biological system when the concentration of MLSS was increased.

Comparative study of the use of pure oxygen and air in the nitrification of a MBR system used for wastewater treatment

A MBR (membrane bioreactor) was performed to quantify the improvement in nitrogen removal terms when pure oxygen and air were used to supply the aerobic conditions at different HRT (hydraulic retention time) and mixed liquid suspended solids concentrations. The improvement in the efficiency was 8.0% and 13.5% when pure oxygen was used with 12h and 18 h of HRT respectively. The ammonium utilization rates with HRT of 18 h with pure oxygen was 1.23 mg N(t)/(L h) and with air 0.66 mg N(t)/(L h) and with HRT of 12h was 1.14 mg N(t)/(L h) and 1.18 mg N(t)/(L h) respectively. Model simulation (DESSAS software) confirmed the validity for nitrogen removal, hydrolysis constant was increased (106% and 184%), decay constant was decreased and Monods constant was increased (190.4% to 324.1%), and a multivariate analysis with CANOCO v4.5 demonstrated that temperature and HRT have the higher influence in kinetics.

Interlinkages between bacterial populations dynamics and the operational parameters in a moving bed membrane bioreactor treating urban sewage.

Bacteria are key players in biological wastewater treatments (WWTs), thus a firm knowledge of the bacterial population dynamics is crucial to understand environmental/operational factors affecting the efficiency and stability of the biological depuration process. Unfortunately, little is known about the microbial ecology of the advanced biological WWTs combining suspended biomass (SB) and attached biofilms (AB). This study explored in depth the bacterial community structure and population dynamics in each biomass fraction from a pilot-scale moving bed membrane bioreactor (MBMBR) treating municipal sewage, by means of temperature-gradient gel electrophoresis (TGGE) and 454-pyrosequencing. Eight experimental phases were conducted, combining different carrier filling ratios, hydraulic retention times and concentrations of mixed liquor total suspended solids. The bacterial community, dominated by Proteobacteria (20.9-53.8%) and Actinobacteria (20.6-57.6%), was very similar in both biomass fractions and able to maintain its functional stability under all the operating conditions, ensuring a successful and steady depuration process. Multivariate statistical analysis demonstrated that solids concentration, carrier filling ratio, temperature and organic matter concentration in the influent were the significant factors explaining population dynamics. Bacterial diversity increased as carrier filling ratio increased (from 20% to 35%, v/v), and solids concentration was the main factor triggering the shifts of the community structure. These findings provide new insights on the influence of operational parameters on the biology of the innovative MBMBRs.

Influence of sludge retention time and temperature on the sludge removal in a submerged membrane bioreactor: Comparative study between pure oxygen and air to supply aerobic conditions

Performance of a bench-scale wastewater treatment plant, which consisted of a membrane bioreactor, was monitored daily using pure oxygen and air to supply aerobic conditions with the aim of studying the increases of the aeration and sludge removal efficiencies and the effect of the temperature. The results showed the capacity of membrane bioreactor systems for removing organic matter. The alpha-factors of the aeration were determined for six different MLSS concentrations in order to understand the system working when pure oxygen and air were used to supply aerobic conditions in the system. Aeration efficiency was increased between 30.7 and 45.9% when pure oxygen was used in the operation conditions (a hydraulic retention time of 12 h and MLSS concentrations between 4,018 and 11,192 mg/L). Sludge removal efficiency increased incrementally, from 0.2 to 1.5% when pure oxygen was used at low sludge retention time and from 1.5% to 15.4% at medium sludge retention time when temperature conditions were lower than 20°C. Moreover, the difference between calculated and experimental sludge retention time was lesser when pure oxygen was used to provide aerobic conditions, so the influence of the temperature decreased when the pure oxygen was used. These results showed the convenience of using pure oxygen due to the improvement in the performance of the system.

Comparative analysis of microbial DNA extraction protocols for groundwater samples

A comparative analysis of four different DNA extraction protocols was performed to determine the best choice for groundwater microbial diversity studies using temperature gradient gel electrophoresis (TGGE) analysis. The methods used were a chelex-based method, a modified salting out procedure (MSOP), and the commercial kits Epicentre and FastDNA. Both commercial kits exhibited the greatest reproducibility in their methods; however, their band patterns were very different. The protocol that showed the highest diversity was the chelex-based method, and the one that showed the lowest diversity was the FastDNA kit.