Assessing the abundance of fungal populations in a full-scale membrane bioreactor (MBR) treating urban wastewater by using quantitative PCR (qPCR).

The abundance of fungi in a full-scale membrane bioreactor (MBR) treating urban wastewater and experiencing seasonal foaming was assessed by quantitative PCR (qPCR), comparing three different sets of widely used universal fungal primers targeting the gene encoding the small ribosomal subunit RNA, 18S-rDNA, (primers NS1-Fung and FungiQuant) or the internal transcribed spacer ITS2 (primers ITS3-ITS4). Fungi were a numerically important fraction of the MBR microbiota (≥106 18S-rDNA copies/L activated sludge), and occurred both in the aerated and anoxic bioreactors. The numbers of copies of fungal markers/L activated sludge calculated using the NS1-Fung or ITS3-ITS4 primer sets were up to 2 orders of magnitude higher than the quantifications based on the FungiQuant primers. Fungal 18S-rDNA counts derived from the FungiQuant primers decreased significantly during cold seasons, concurring with foaming episodes in the MBR. Redundancy analysis corroborated that temperature was the main factor driving fungi abundance, which was also favored by longer solid retention time (SRT), lower chemical oxygen demand/biochemical oxygen demand at 5 days (COD/BOD5) of influent water, and lower biomass accumulation in the MBR.

Prevalence of Nitrosomonas cluster 7 populations in the ammonia-oxidizing community of a submerged membrane bioreactor treating urban wastewater under different operation conditions.

A pilot-scale ultrafiltration membrane bioreactor (MBR) was used for the aerobic treatment of urban wastewater in four experimental stages influenced by seasonal temperature and different sets of operation conditions. The structure of the ammonia-oxidizing bacteria (AOB) community was profiled by temperature gradient gel electrophoresis (TGGE), based on the amplification and separation of partial ammonia-monoxygenase subunit A (amoA) genes. Canonical correspondence analysis revealed that temperature, hydraulic retention time and percentage of ammonia removal had a significant effect on the fingerprints of AOB communities. Phylogenetic analysis conducted on amoA/AmoA sequences of reamplified TGGE bands showed, however, that closely related ammonia-oxidizing populations inhabited the sludge of the MBR in all experimental stages. Nitrosomonas cluster 7 populations (N. europaea-N. eutropha cluster) prevailed under all conditions tested, even when the MBR was operated under complete biomass retention or at low temperatures, suggesting that the high ammonia concentrations in the system were determinant to select r-strategist AOB.

Microbial indicators of efficient performance in an anaerobic/anoxic/aerobic integrated fixed-film activated sludge (A2O-IFAS) and a two-stage mesophilic anaerobic digestion process.

An analysis of the community structure, diversity and population dynamics of Bacteria and Archaea in the suspended and attached biomass fractions of a pilot-scale anaerobic/anoxic/aerobic integrated fixed-film activated sludge (A2O-IFAS) was executed. Along with this, the effluents of the acidogenic (AcD) and methanogenic (MD) digesters of a two-stage mesophilic anaerobic (MAD) system treating the primary sludge (PS) and waste activated sludge (WAS) generated by the A2O-IFAS were also analyzed. Non-metric multidimensional scaling (MDS) and Biota-environment (BIO-ENV) multivariate analyses were performed to link population dynamics of Bacteria and Archaea to operating parameters and removal efficiencies of organic matter and nutrients, in search of microbial indicators associated with optimal performance. In all samples analyzed, Proteobacteria, Bacteroidetes and Chloroflexi were the most abundant phyla, while the hydrogenotrophic methanogens Methanolinea, Methanocorpusculum and Methanobacterium were the predominant archaeal genera. BIO-ENV analysis disclosed strong correlations between the population shifts observed in the suspended and attached bacterial communities of the A2O-IFAS and the removal rates of organic matter, N and P. It is noteworthy that the incorporation of carriers combined with a short sludge retention time (SRT = 4.0 ± 1.0 days) enhanced N removal performance of the A2O by favoring the enrichment of bacterial genera able to denitrify (Bosea, Dechloromonas, Devosia, Hyphomicrobium, Rhodobacter, Rhodoplanes, Rubrivivax, and Sulfuritalea) in the attached biomass fraction. In addition, operation at short SRT enabled the generation of a highly biodegradable WAS, which enhanced the biogas and methane yields in the two-stage MAD. An increase in the relative abundance of Acetobacteroides (uncultured Blvii28 wastewater-sludge group of Rikenellaceae family) correlated positively with the volatile solids removal rate (%VSR), CH4 recovery rate and %CH4 in the biogas (r > 0.8), supporting their relevance for an efficient methanogenesis in two-stage systems.

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.

Submerged filter biofilm formation by nitrate-contaminated groundwater microbiota.

Denitrifying biofilms developed in a lab-scale submerged filter by autochthonous bacteria from nitrate-contaminated groundwater were studied. The system was supplied with groundwater (16 mg N-NO(3)(-)/L), from which the oxygen had been eliminated and to which an excess of carbon source had been added. The reactor was incubated in a thermostated chamber at 5°C, 10°C, 20°C and 30°C. Colonization of the support was studied using surface scanning microscopy, and biofilm bacterial composition was studied by PCR/TGGE. Support material was colonized at all the temperatures assayed, although this parameter affected the growth of the biofilm, which developed most at temperatures over 20°C. The composition of bacterial communities varied according to the temperature. Community profiles of the biofilm formed at 5°C and 10°C clustered away from those of the biofilm formed at 20°C and 30°C. 16S rDNA sequences reveled that the biofilm was mainly composed of psychrotolerant species typically inhabiting freshwaters, and we obtained sequencing bands that were affiliated to denitrifying and non-denitrifying heterotrophic species. The extent of colonization was low when compared to previously inoculated systems, and the capacity for nitrate elimination was also low.

Insights into the removal of pharmaceutically active compounds from sewage sludge by two-stage mesophilic anaerobic digestion.

The removal efficiencies (REs) of twenty-seven pharmaceutically active compounds (PhACs) (eight analgesic/anti-inflammatories, six antibiotics, four ß-blockers, two antihypertensives/diuretics, three lipid regulators and four psychiatric drugs) were evaluated in a pilot-scale two-stage mesophilic anaerobic digestion (MAD) system treating thickened sewage sludge from a pilot-scale A2O™ wastewater treatment plant (WWTP) which was fed with wastewater from the pre-treatment of the full-scale WWTP Murcia Este (Murcia, Spain). The MAD system was long-term operated using two different sets of sludge retention times (SRTs) for the acidogenic (AcD) and methanogenic (MD) digesters (phase I, 2 and 12 days; and phase II, 5 and 24 days, in AcD and MD, respectively). Quantitative PCR (qPCR) and Illumina MiSeq sequencing were used to estimate the absolute abundance of Bacteria, Archaea, and Fungi and investigate the structure, diversity and population dynamics of their communities in the AcD and MD effluents. The extension of the SRT from 12 (phase I) to 24 days (phase II) in the MD was significantly linked with an improved removal of carbamazepine, clarithromycin, codeine, gemfibrozil, ibuprofen, lorazepam, and propranolol. The absolute abundances of total Bacteria and Archaea were higher in the MD regardless of the phase, while the diversity of bacterial and archaeal communities was lower in phase II, in both digesters. Non-metric multidimensional scaling (MDS) plots showed strong negative correlations among phyla Proteobacteria and Firmicutes and between genera Methanosaeta and Methanosarcina throughout the full experimental period. Strong positive correlations were revealed between the relative abundances of Methanospirillum and Methanoculleus and the methanogenesis performance parameters (volatile solids removal, CH4 recovery rate and %CH4 in the biogas), which were also related to longer SRT. The REs of several PhACs (naproxen, ketoprofen, ofloxacin, fenofibrate, trimethoprim, and atenolol) correlated positively (r > 0.75) with the relative abundances of specific bacterial and archaeal groups, suggesting their participation in biodegradation/biotransformation pathways.

Linking microbial diversity and population dynamics to the removal efficiency of pharmaceutically active compounds (PhACs) in an anaerobic/anoxic/aerobic (A2O) system.

The removal efficiencies (REs) of nineteen pharmaceutically active compounds (PhACs) (six antibiotics-clarithromycin, ofloxacin, sulfadiazine, sulfamethazine, sulfamethoxazole and trimethoprim -, four ß-blockers -atenolol, metoprolol, propranolol and sotalol-, two antihypertensives/diuretics -furosemide and hydrochlorothiazide-, three lipid regulators -bezafibrate, fenofibrate and gemfibrozil-, and four psychiatric medications -carbamazepine, diazepam, lorazepam and paroxetine) were ascertained in a pilot-scale anaerobic/anoxic/aerobic (A2O) system treating urban wastewater, long term operated during two experimental phases using different sets of environmental conditions and operating parameters. Illumina MiSeq sequencing was used to investigate the structure, diversity and population dynamics of bacteria, archaea and fungi communities in the activated sludge. The results showed that mixed liquor suspended solids (MLSS) and food-to-microorganisms ratio (F/M) were operational parameters significantly influencing the REs of five of the analyzed PhACs in the A2O system. Biota-environment (BIO-ENV) analysis revealed strong correlations between population shifts of the activated sludge community and the REs of PhACs of the different pharmaceutical families. Increased REs of clarithromycin, furosemide, bezafibrate and gemfibrozil were concomitant to higher relative abundances of bacterial phylotypes classified within the Rhodobacteraceae and Sphingomonadaceae (Alphaproteobacteria), while those of Betaproteobacteria, Chloroflexi and Methanomethylovorans (Euryarchaea) correlated positively with the REs of up to seven PhACs belonging to different therapeutic groups.

Abundance of total and metabolically active Candidatus Microthrix and fungal populations in three full-scale wastewater treatment plants.

The abundances of total and metabolically active populations of Candidatus Microthrix and Fungi were evaluated by quantitative PCR (qPCR) and retrotranscribed qPCR of ribosomal molecular markers in three different full-scale wastewater treatment plants (WWTPs), in absence of bulking/foaming episodes. Significant differences of the abundance of rDNAs and rRNAs of Candidatus Microthrix and Fungi were observed among the three WWTPs. The average relative abundances of 16S rDNA copies of Candidatus Microthrix to those of Bacteria ranged 3.4-8.9%. Biota-environment analysis (BIO-ENV) demonstrated that the number of copies of both 16S rDNA and rRNA of Candidatus Microthrix increased at longer hydraulic and solids' retention times and with higher nitrate concentrations in the activated sludge. The abundance of Candidatus Microthrix correlated strongly and positively with the removal efficiencies of organic matter and total nitrogen in the tested WWTPs, highlighting the role of these particular microbial group in the performance of these engineered systems.

Removal of anti-inflammatory/analgesic pharmaceuticals from urban wastewater in a pilot-scale A2O system: Linking performance and microbial population dynamics to operating variables.

In this study, the removal rates of eight anti-inflammatory and/or analgesic pharmaceuticals, AIAPs (acetaminophen, ibuprofen, naproxen, ketoprofen, diclofenac, codeine, indomethacin and propyphenazone) were assessed in a pilot-scale A2O system (including anaerobic/anoxic/aerobic zones), long term operated during two experimental phases using different sets of environmental conditions and operating parameters. qPCR was used to quantify the absolute abundances of total Bacteria, total Archaea, mycolic-acid containing filamentous Actinobacteria (Mycolata) and Fungi within the activated sludge microbial community developed in the system. Multivariate analyses and Spearman correlation coefficients were used in search of significant links among the removal rates of the AIAPs, the abundances of the targeted microbial groups in the activated sludge, and the changes of environmental/operating variables in the A2O system. Improved removal efficiencies of several of the AIAPs analyzed (acetaminophen, ibuprofen, naproxen, ketoprofen) were correlated to higher organic load in the influent water, higher concentration of mixed liquor suspended solids (MLSS), lower temperature and lower food-to-microorganisms ratio (F/M). Removal efficiencies of several pharmaceuticals correlated with increased abundances of Mycolata in the A2O system, pointing at this group of bacteria as candidate key players for AIAPs removal in activated sludge.

Linking nitrous oxide emissions to population dynamics of nitrifying and denitrifying prokaryotes in four full-scale wastewater treatment plants.

Ammonia-oxidizing bacteria (AOB), ammonia oxidizing archaea (AOA) and N2O-reducing denitrifiers were measured by quantitative real-time PCR (qPCR) in activated sludge samples from four full-scale wastewater treatment plants (WWTPs) in South Spain, and their abundances were linked to the generation of N2O in the samples using multivariate analysis (Non-metric multidimensional scaling, MDS, and BIO-ENV). The average abundances of AOA remained in similar orders of magnitude in all WWTPs (106 copies amoA/L activated sludge mixed liquor), while significant differences were detected for AOB (105-109copies amoA/L) and N2O-reducers (107-1010copies nosZ/L). Average N2O emissions measured in activated sludge samples ranged from 0.10 ±â€¯0.05 to 6.49 ±â€¯8.89 mg N2O-N/h/L activated sludge, and were strongly correlated with increased abundances of AOB and lower counts of N2O-reducers. A significant contribution of AOA to N2O generation was unlikely, since their abundance correlated negatively to N2O emissions. AOB abundance was favoured by higher NO3- and NO2-concentrations in the activated sludge.

Influence of temperature on microbial colonisation of clayey schist as a support media of a submerged filter for groundwater denitrification.

The effect of temperature on biofilm formation and denitrification activity was evaluated. Assays were made in a lab-scale submerged filter for the denitrification of polluted groundwater, with and without a previous inoculation. The inoculation was carried out with a selected strain of Pseudomonas mandelii. Different temperatures were tested: 5, 10, 20 and 30 degrees C. Biofilm observations were made, and monitoring of the denitrification capacity of the system was maintained during the experiment. Our results showed that both colonisation of the support material of the filter and biofilm maturity have a dependency with temperature, with an optimum temperature of 20 'C if the system was previously inoculated with the Pseudomonas mandelii strain. For a correct achievement of the denitrification process, a previous inoculation of the system is essential. Although the development of a biofilm from the natural microbiota present in the groundwater is possible, it is not capable to adequately denitrify polluted groundwater. In terms of the correct achievement of the denitrification process, temperature affects the operation of the system at cold environments, although the use of Pseudomonas mandelii strain A103 allows denitrification at 10-30 degrees C with very good results (above 90% removal), affecting only to the time needed for the stabilisation of the system.

Full-scale photobioreactor for biotreatment of olive washing water: Structure and diversity of the microalgae-bacteria consortium.

The performance of a full-scale photobioreactor (PBR) for the treatment of olive washing water (OWW) was evaluated under different HRTs (5-2days). The system was able to treat up to 3926L OWWday-1, and consisted of an activated-carbon pretreatment column and a tubular PBR unit (80 tubes, 98.17L volume, 2-m height, 0.25m diameter). PBR was an effective and environmentally friendly method for the removal of phenols, COD, BOD5, turbidity and color from OWW (average efficiencies 94.84±0.55%, 85.86±1.24%, 99.12±0.17%, 95.86±0.98% and 87.24±0.91%, respectively). The diversity of total bacteria and microalgae in the PBR was analyzed using Illumina-sequencing, evaluating the efficiency of two DNA extraction methods. A stable microalgae-bacteria consortium was developed throughout the whole experimentation period, regardless of changes in HRT, temperature or solar radiation. MDS analyses revealed that the interplay between green algae (Sphaeropleales), cyanobacteria (Hapalosiphon) and Proteobacteria (Rhodopseudomonas, Azotobacter) played important roles in OWW bioremediation.

Selection and identification of bacteria isolated from waste crude oil with polycyclic aromatic hydrocarbons removal capacities.

Fifteen bacterial strains isolated from solid waste oil samples were selected due to their capacity of growing in the presence of hydrocarbons. The isolates were identified by PCR of the 16S rDNA gene using fD1 and rD1 primers. The majority of the strains belonged to genera Bacillus, Bacillus pumilus (eight strains) and Bacillus subtilis (two strains). Besides, three strains were identified as Micrococcus luteus, one as Alcaligenes faecalis and one strain as Enterobacter sp. Growth of the above-mentioned strains in mineral liquid media amended with naphthalene, phenanthrene, fluoranthene or pyrene as sole carbon source was studied and our results showed that these strains can tolerate and remove different polycyclic aromatic hydrocarbons that may be toxic in the environment polluted with hydrocarbons. Finally, the capacity of certain strains to emulsify octane, xilene, toluene, mineral oil and crude oil, and its ability to remove hydrocarbons, look promising for its application in bioremediation technologies.

Community structure, population dynamics and diversity of fungi in a full-scale membrane bioreactor (MBR) for urban wastewater treatment.

Community structure, population dynamics and diversity of fungi were monitored in a full-scale membrane bioreactor (MBR) operated throughout four experimental phases (Summer 2009, Autumn 2009, Summer 2010 and Winter, 2012) under different conditions, using the 18S-rRNA gene and the intergenic transcribed spacer (ITS2-region) as molecular markers, and a combination of temperature-gradient gel electrophoresis and 454-pyrosequencing. Both total and metabolically-active fungal populations were fingerprinted, by amplification of molecular markers from community DNA and retrotranscribed RNA, respectively. Fingerprinting and 454-pyrosequencing evidenced that the MBR sheltered a dynamic fungal community composed of a low number of species, in accordance with the knowledge of fungal diversity in freshwater environments, and displaying a medium-high level of functional organization with few numerically dominant phylotypes. Population shifts were experienced in strong correlation with the changes of environmental variables and operation parameters, with pH contributing the highest level of explanation. Phylotypes assigned to nine different fungal Phyla were detected, although the community was mainly composed of Ascomycota, Basidiomycota and Chytridiomycota/Blastocladiomycota. Prevailing fungal phylotypes were affiliated to Saccharomycetes and Chytridiomycetes/Blastocladiomycetes, which displayed antagonistic trends in their relative abundance throughout the experimental period. Fungi identified in the activated sludge were closely related to genera of relevance for the degradation of organic matter and trace-organic contaminants, as well as genera of dimorphic fungi potentially able to produce plant operational issues such as foaming or biofouling. Phylotypes closely related to genera of human and plant pathogenic fungi were also detected.

The ratio of metabolically active versus total Mycolata populations triggers foaming in a membrane bioreactor.

The abundance of total and metabolically active populations of Mycolata was evaluated in a full-scale membrane bioreactor (MBR) experiencing seasonal foaming, using quantitative PCR (qPCR) and retrotranscribed qPCR (RT-qPCR) targeting the 16S rRNA gene sequence. While the abundance of total Mycolata remained stable (10(10) copies of 16S rRNA genes/L activated sludge) throughout four different experimental phases, significant variations (up to one order of magnitude) were observed when the 16S rRNA was targeted. The highest ratios of metabolically active versus total Mycolata populations were observed in samples of two experimental phases when foaming was experienced in the MBR. Non-metric multidimensional scaling and BIO-ENV analyses demonstrated that this ratio was positively correlated to the concentrations of substrates in the influent water, F/M ratio, and pH, and negatively correlated to temperature and solids retention time. It the first time that the ratio of metabolically active versus total Mycolata is found to be a key parameter triggering foaming in the MBR; thus, we propose it as a candidate predictive tool.

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.

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.

Effects of culture conditions on the production of polyhydroxyalkanoates by Azotobacter chroococcum H23 in media containing a high concentration of alpechín (wastewater from olive oil mills) as primary carbon source.

Large amounts of homopolymers containing beta-hydroxybutyrate (PHB) and copolymers containing beta-hydroxyvalerate (P[HB-co-HV]) are produced by Azotobacter chroococcum strain H23 when growing in culture media amended with alpechín (wastewater from olive oil mills) as the sole carbon source. Copolymer was formed when valerate (pentanoate) was added as a precursor to the alpechín medium, but it was not formed with the addition of propionate as a precursor. A. chroococcum formed homo- and copolymers of polyhydroxyalkanoates (PHAs) up to 80% of the cell dry weight, when grown on NH(4)(+)-medium supplemented with 60% (v/v) alpechín, after 48 h of incubation at 100 rev min(-1) and 30 degrees C. Production of PHAs by strain H23 using alpechín looks promising, as the use of a cheap substrate for the production of these materials is essential if bioplastics are to become competitive products.

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.

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.