J.M. Poyatos

Comparative analysis of the bacterial diversity in a lab-scale moving bed biofilm reactor (MBBR) applied to treat urban wastewater under different operational conditions

Different types of carriers were tested as support material in a lab-scale moving bed biofilm reactor (MBBR) used to treat urban wastewater under three different conditions of hydraulic retention time (HRT) and carrier filling ratios (FR). The bacterial diversity developed on the biofilms responsible of the treatment was studied using a cultivation-independent approach based on the polymerase chain reaction-temperature gradient gel electrophoresis technique (PCR-TGGE). Cluster analysis of TGGE fingerprints showed significant differences of community structure dependent upon the different operational conditions applied. Redundancy analysis (RDA) was used to determine the relationship between the operational conditions (type of carrier, HRT, FR) and bacterial biofilm diversity, demonstrating a significant effect of FR=50%. Phylogenetic analysis of PCR-reamplified and sequenced TGGE bands revealed that the prevalent Bacteria populations in the biofilm were related to Betaproteobacteria (46%), Firmicutes (34%),Alphaproteobacteria (14%) and Gammaproteobacteria (9%).

Microbial community structure and dynamics in a pilot-scale submerged membrane bioreactor aerobically treating domestic wastewater under real operation conditions.

A pilot scale submerged ultra-filtration membrane bioreactor (MBR) was used for the aerobic treatment of domestic wastewater over 9 months of year 2006 (28th March to 21st December). The MBR was installed at a municipal wastewater facility (EMASAGRA, Granada, Spain) and was fed with real wastewater. The experimental work was divided in 4 stages run under different sets of operation conditions. Operation parameters (total and volatile suspended solids, dissolved oxygen concentration) and environmental variables (temperature, pH, COD and BOD(5) of influent water) were daily monitored. In all the experiments conducted, the MBR generated an effluent of optimal quality complying with the requirements of the European Law (91/271/CEE 1991). A cultivation-independent approach (polymerase chain reaction-temperature gradient gel electrophoresis, PCR-TGGE) was used to analyze changes in the structure of the bacterial communities in the sludge. Cluster analysis of TGGE profiles demonstrated significant differences in community structure related to variations of the operation parameters and environmental factors. Canonical correspondence analysis (CCA) suggested that temperature, hydraulic retention time and concentration of volatile suspended solids were the factors mostly influencing community structure. 23 prominent TGGE bands were successfully reamplified and sequenced, allowing gaining insight into the identities of predominantly present bacterial populations in the sludge. Retrieved partial 16S-rRNA gene sequences were mostly related to the alpha-Proteobacteria, beta-Proteobacteria and gamma-Proteobacteria classes. The community established in the MBR in each of the four stages of operation significantly differed in species composition and the sludge generated displayed dissimilar rates of mineralization, but these differences did not influence the performance of the bioreactor (quality of the permeate). These data indicate that the flexibility of the bacterial community in the sludge and its ability to get adapted to environmental changes play an important role for the stable performance of MBRs.

Microbial enzymatic activities in a pilot-scale MBR experimental plant under different working conditions.

Phosphatases, glucosidase, protease, esterase and dehydrogenase activities in a MBR (membrane bioreactor) system equipped with ultrafiltration membranes for the treatment of real urban wastewater were measured at different volatile suspended solid (VSS) concentrations, total suspended solid (TSS) concentrations, hydraulic retention times (HRT), temperatures and inflow rates. The results showed the capacity of the MBR system to remove COD and BOD(5) at TSS between 7200 and 13,300 mg/L; HRT values of 8.05 and 15.27 h; inflow rates of 14.67 and 27.81 L/h; and temperatures between 4 and 27 degrees C. The enzymatic activities are influenced by increases in VSS and TSS concentrations. These results suggest that the ability to get adapted to environmental changes of the bacterial populations and their microbial enzymatic activities is essential to understand the biological processes that occur in MBR systems and crucial for proper urban wastewater treatment when using MBR technologies.

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%).

Structure of archaeal communities in membrane-bioreactor and submerged-biofilter wastewater treatment plants.

A cultivation independent approach (PCR-TGGE) was used to evaluate the occurrence of Archaea in four wastewater treatments based on technologies other than activated sludge, and to comparatively analyze their community structure. TGGE fingerprints (based on partial archaeal 16S-rRNA amplicons) were obtained from sludge samples taken from a pilot-scale aerated MBR fed with urban wastewater and operated under two different sets of conditions (MBR1 and MBR2 treatments), and also from biofilms sampled from two pilot-scale submerged biofilters (SBs) consisting of one aerated and one anoxic column each, fed with urban (USB treatment) or industrial (ISB treatment) wastewater, respectively. Analysis of TGGE fingerprints revealed clear and significant differences of the community structure of Archaea between the wastewater treatments studied, primarily according to wastewater origin and the type of technology. Thirty-two different band classes were detected among the 23 sludge and biofilm samples analyzed, from which five were selected as dominant or distinctive of the four treatments studied. Sixteen predominant TGGE bands were identified, revealing that all of them were related to methanogenic Archaea. Neither other Euryarchaeota groups nor Crenarchaeota members were identified amongst the 16S-rRNA fragments sequenced from separated TGGE bands.

Effect of the mixed liquor suspended solid on permeate in a membrane bioreactor system applied for the treatment of sewage mixed with wastewater of the milk from the dairy industry.

The performance of a bench-scale submerged membrane bioreactor (MBR) equipped with ultrafiltration membranes (ZENON) was investigated at different mixed liquor suspended solid (MLSS) concentrations (3069, 4314 and 6204 mg/L). The pilot plant was located in the wastewater treatment plant of the city of Granada (Puente de los Vados, Granada, Spain), which receives the wastewater of the milk from the dairy industry of Granada. The results showed the capacity of the MBR systems to remove organic material (COD and BOD5), suspended solids, turbidity, color and microbial indicators such as E. coli and coliphages. Therefore, the results suggest that the transmembrane pressure (TMP) was influence by the MLSS concentration assayed. However, an increase in the MLSS concentration increases the nitrification processes and consequently the amount of NO3 − in permeate.

Water quality characterization in real biofilm wastewater treatment systems by particle size distribution.

This article analyses the relation between wastewater quality parameters and particle size distribution (PSD) in three real wastewater treatment plants with different biofilm technologies: submerged biofilter system, trickling filter system, and rotating biological contactor system. The main quality parameters, (suspended solids, turbidity and COD), and PSD in the influent and effluent water of each different biofilm treatment were analyzed during 1year. The PSD was fitted using the power law (n(d(p))= partial differentialN(d(p))/ partial differentiald(p)=A x d(p)(-)(b)(Log(d)(p)())) obtaining the coefficients A and b to define the particle distribution. Mathematical correlations between this coefficients and the rest of parameters studied were found (SS=0.0713 x A(0.585), turbidity=4.549 x 10(-4) x A(1.096), COD=0.0201 x A(0.774)). The relation with the average particle size by mass was also found, (d(pma)=60.3137 x b(-2.242)). Moreover a relation between PSD and the particle elimination efficiency of the secondary treatment was study, (eta=2.844-2.498 x b(relative)+0.0863 x A(relative)). These expressions are very useful to understand the behavior of the biofilm treatment system using PSD analyses.

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.

Moving bed biofilm reactor to treat wastewater

This review carries out a comparative study of advanced technologies to design, upgrade and rehabilitate wastewater treatment plants. The study analyzed the relevant researches in the last years about the moving bed biofilm reactor process with only attached biomass and with hybrid biomass, which combined attached and suspended growth; both could be coupled with a secondary settling tank or microfiltration/ultrafiltration membrane as a separation system. The physical process of membrane separation improved the organic matter and NH4+-N removal efficiencies compared with the settling tank. In particular, the pure moving bed biofilm reactor–membrane bioreactor showed average chemical oxygen demand, biochemical oxygen demand on the fifth day and total nitrogen removal efficiencies of 88.32, 90.84 and 60.17%, respectively, and the hybrid moving bed biofilm reactor–membrane bioreactor had mean chemical oxygen demand, biochemical oxygen demand on the fifth day and total nitrogen reduction percentages of 91.18, 97.34 and 68.71%, respectively. Moreover, the hybrid moving bed biofilm reactor–membrane bioreactor showed the best efficiency regarding organic matter removal for low hydraulic retention times, so this system would enable the rehabilitation of activated sludge plants and membrane bioreactors that did not comply with legislation regarding organic matter removal. As the pure moving bed biofilm reactor–membrane bioreactor performed better than the hybrid moving bed biofilm reactor–membrane bioreactor concerning the total nitrogen removal under low hydraulic retention times, this system could be used to adapt wastewater treatment plants whose effluent was flowed into sensitive zones where total nitrogen concentration was restricted. This technology has been reliably used to upgrade overloaded existing conventional activated sludge plants, to treat wastewater coming from textile, petrochemical, pharmaceutical, paper mill or hospital effluents, to treat wastewater containing recalcitrant compounds efficiently, and to treat wastewater with high salinity and/or low and high temperatures.

Isolation and metagenomic characterization of bacteria associated with calcium carbonate and struvite precipitation in a pure moving bed biofilm reactor-membrane bioreactor

A bench-scale pure moving bed bioreactor-membrane bioreactor (MBBR-MBR) used for the treatment of urban wastewater was analyzed for the identification of bacterial strains with the potential capacity for calcium carbonate and struvite biomineral formation. Isolation of mineral-forming strains on calcium carbonate and struvite media revealed six major colonies with a carbonate or struvite precipitation capacity in the biofouling on the membrane surface and showed that heterotrophic bacteria with the ability to precipitate calcium carbonate and struvite constituted ~7.5% of the total platable bacteria. These belonged to the genera Lysinibacillus, Trichococcus, Comamomas and Bacillus. Pyrosequencing analysis of the microbial communities in the suspended cells and membrane biofouling showed a high degree of similarity in all the samples collected with respect to bacterial assemblage. The study of operational taxonomic units (OTUs) identified through pyrosequencing suggested that ~21% of the total bacterial community identified in the biofouling could potentially form calcium carbonate or struvite crystals in the pure MBBR-MBR system used for the treatment of urban wastewater.