Linking operation parameters and environmental variables to population dynamics of Mycolata in a membrane bioreactor.

The community structure and population dynamics of Mycolata were monitored in a full-scale membrane bioreactor during four experimental phases under changing operating and environmental conditions, by means of temperature-gradient gel electrophoresis of partial 16S-rRNA genes amplified from community DNA and RNA templates (total and active populations). Non-metric multidimensional scaling and BIO-ENV analyses demonstrated that population dynamics were mostly explained (30-32%) by changes in the input of nutrients in the influent water and the accumulation of biomass in the bioreactors, while the influence of hydraulic and solid retention times, temperature and F/M ratio was minor. Significant correlations were observed between particular Mycolata phylotypes and one or more variables, contributing information for the prediction of their abundance and activity under changing conditions. Fingerprinting and multivariate analyses demonstrated that two foaming episodes, recorded at temperatures <20°C, were connected to the increase of the relative abundance of Mycolata unrelated to Gordonia amarae.

Quantitative response of nitrifying and denitrifying communities to environmental variables in a full-scale membrane bioreactor.

The abundance and transcription levels of specific gene markers of total bacteria, ammonia-oxidizing Betaproteobacteria, nitrite-oxidizing bacteria (Nitrospira-like) and denitrifiers (N2O-reducers) were analyzed using quantitative PCR (qPCR) and reverse-transcription qPCR during 9 months in a full-scale membrane bioreactor treating urban wastewater. A stable community of N-removal key players was developed; however, the abundance of active populations experienced sharper shifts, demonstrating their fast adaptation to changing conditions. Despite constituting a small percentage of the total bacterial community, the larger abundances of active populations of nitrifiers explained the high N-removal accomplished by the MBR. Multivariate analyses revealed that temperature, accumulation of volatile suspended solids in the sludge, BOD5, NH4(+) concentration and C/N ratio of the wastewater contributed significantly (23-38%) to explain changes in the abundance of nitrifiers and denitrifiers. However, each targeted group showed different responses to shifts in these parameters, evidencing the complexity of the balance among them for successful biological N-removal.

Exploring the links between population dynamics of total and active bacteria and the variables influencing a full-scale membrane bioreactor (MBR).

Long-term dynamics of total and active bacterial populations in a full-scale membrane bioreactor (MBR) treating urban wastewater were monitored during nine months by temperature-gradient gel electrophoresis (TGGE) of partial 16S-rRNA genes, amplified from community DNA and RNA templates. The bacterial community, dominated by Alphaproteobacteria, displayed the required characteristics for a successful and steady contaminant removal under real operating conditions. The evolution of population dynamics showed that a fully-stable microbial community was not developed even after technical stabilization and steady performance of the MBR were achieved. Non-metric multidimensional scaling and BIO-ENV demonstrated that the trends of the populations were often mostly explained by temperature, followed by the concentration of volatile suspended solids and C/N ratio of the influent. These variables were mainly responsible for triggering the shifts between functionally redundant populations. These conclusions contribute to the prediction of the complex profiles of adaptation and response of bacterial populations under changing conditions.

Linking hydrolytic activities to variables influencing a submerged membrane bioreactor (MBR) treating urban wastewater under real operating conditions.

The seasonal variation of the hydrolytic activities acid and alkaline phosphatase, α-glucosidase and protease, was studied in both the aerated and anoxic phases of a full-scale membrane bioreactor (MBR) (total operational volume = 28.2 m(3)), operated in pre-denitrification mode and fed real urban wastewater. Non-metric multidimensional scaling (MDS) and BIO-ENV analysis were used to study the distribution of enzyme activities in different seasons of the year (spring, summer and autumn) and unveil their relationships with changes in variables influencing the system (composition of influent wastewater, activated sludge temperature and biomass concentration in the bioreactors). The activities of all the tested hydrolases were remarkably dynamic, and each enzyme showed complex and diverse patterns of variation. Except in the summer season, the variables included in this study gave a good explanation of those patterns and displayed high and consistent correlations with them; however, markedly different correlation trends were found in each season, indicating dissimilar adaptation responses of the community to the influence of changing conditions. A consistent and highly negative correlation between protease and α-glucosidase was revealed in all the experiments. The variables included in this study showed contrary influences on these activities, suggesting an alternation of the major groups of carbon-degrading hydrolases in connection to changes in temperature and the availability and composition of nutrients in the different seasons. Sampling over a long period of time was required to adequately lay down the links between hydrolytic activities and the variables influencing the MBR system. These results highlight the complexity of the regulation of substrate degradation by the mixed microbial sludge communities under real operating conditions.

Characterization of bacterial communities exposed to Cr(III) and Pb(II) in submerged fixed-bed biofilms for groundwater treatment.

Two pilot-scale submerged-bed microbial biofilms were set up for the removal of Cr(III) and Pb(II) from groundwater, and the biological activities and structure of the bacterial communities developed in the presence of the heavy metals were analyzed. Artesian groundwater was polluted with Cr(III) or Pb(II) (15 mg/l) and amended with sucrose (250 mg/l) as carbon source. While Pb(II) was over 99% removed from groundwater during long-term operation (130 days), the efficiency of the removal of Cr(III) significantly decreased in time (95-73% after 60 days). Cr(III)-amended biofilms displayed significant lower sucrose consumption, ATP cell contents and alkaline phosphatase activity, compared to biofilms formed in the presence of Pb(II), while analysis of exopolymers demonstrated significant differences in their composition (content of carbohydrates and acetyl groups) in response to each heavy metal. According to transmission electron microscopy (TEM) and electron-dispersive X-ray analysis (EDX), Cr(III) bioaccumulated in the exopolymeric matrix without entering bacterial cells, while Pb(II) was detected both extra and intracellularly, associated to P and Si. Temperature-gradient gel electrophoresis (TGGE) profiling based on partial amplification of 16S rRNA genes was used to analyze the differences in the structure of the biofilm bacterial communities developed under exposure to each heavy metal. Prevalent populations in the biofilms were further identified by reamplification and sequencing of isolated TGGE bands. 75% of the sequences in the Pb(II) biofilter were evolutively close to the Rhodobacterales, while in the Cr(III) biofilter 43% of the sequences were found affiliated to the Rhizobiales and Sphingomonadales, and 57% to Betaproteobacteria.

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.