P. Maza-Márquez

Start-up and operation of an aerobic granular sludge system under low working temperature inoculated with cold-adapted activated sludge from Finland

An aerobic granular sludge system has been started-up and operated at 7°C temperature using cold-adapted activated sludge as inoculum. The system could form granular biomass due to batch operation allowing for just 5-3min of biomass sedimentation. Scanning electron microscopy showed that fungi helped in the granular biomass formation in the early stages of the granule formation. The removal performance of the system was of 92-95% in BOD5, 75-80% in COD, 70-76% in total nitrogen and 50-60% in total phosphorous. The bacterial community structure from cold-adapted activated sludge changed during the operational time, leading to a final configuration dominated by Microbacteriaceae members Microbacterium and Leucobacter, which were strongly correlated to biomass settling velocity and bioreactor performance, as suggested by multivariate redundancy analyses. This experiment showed that aerobic granular sludge systems could be successfully started-up and operated, with high performance, under low operational temperatures when using cold-adapted biomass as inoculum.

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.

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.

Distribution and microbial community structure analysis of a single-stage partial nitritation/anammox granular sludge bioreactor operating at low temperature

ABSTRACT In the last decade, autotrophic nitrogen removal technologies based on anammox metabolism have become state of the art in urban and industrial wastewater treatment systems, due to their advantages over traditional nitrogen removal processes. However, their application is currently limited to the treatment of warm wastewater (25–40°C) mainly due to the low growth rate of the anammox bacteria. The extension of the application field to wastewater characterized by lower temperatures (8–20°C), such as those typical for municipal sewage, allows the design of treatment systems with a net energy production. In this study, the distribution and bacterial community structure of a lab-scale single-stage partial nitritation/anammox (PN/A) granular sludge bioreactor operating at low temperatures was analysed using next-generation sequencing techniques. The presence of ammonium-oxidizing bacteria and anammox bacteria was found, but the appearance of other bacterial species shows a complex microbial ecosystem. Evaluation of ecological roles of representative species inside the single-stage PN/A bioreactor was accomplished. Results obtained will be helpful for the future design and operation of PN/A systems performing at low temperatures.

Performance and microbial community structure of a polar Arctic Circle aerobic granular sludge system operating at low temperature

The aim of this work was to study the performance and microbial community structure of a polar Arctic Circle aerobic granular sludge (AGS) system operating at low temperature. Thus, an AGS bioreactor was operated at 7, 5 and 3 °C of temperature using a cold-adapted sludge from Lapland. At 5 °C, it yielded acceptable conversion rates, in terms of nitrogen, phosphorous, and organic matter. However, under 3 °C a negligible nitrogen and phosphorous removal performance was observed. Below 5 °C, scanning electron microscopy studies showed a wispy, non-dense and irregular granular structure with a strong outgrowth of filamentous. Moreover, Illumina next-generation sequencing showed a heterogeneous microbial population where SM1K20 (Archaea), Trichosporon domesticum (Fungus), and Zooglea, Arcobacter and Acinetobacter (Bacteria) were the dominant phylotypes. Our study suggests that AGS technologies inoculated with North Pole sludge could be operated, in cold regions for a period longer than 3 months (winter season) under 5 °C of water temperature.

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.

Process performance and bacterial community dynamics of partial-nitritation biofilters subjected to different concentrations of cysteine amino acid

Partial‐nitritation processes are used for the biological treatment of high nitrogen‐low organic carbon effluents, such as anaerobic digestion reject water. The release of certain products generated during the anaerobic digestion process, such as amino acids, could potentially reduce the performance of these partial‐nitritation bioprocesses. To investigate this, four partial‐nitritation biofilters were subjected to continuous addition of 0, 150, 300, and 500 mg L−1 cysteine amino acid in their influents. The addition of the amino acid had an impact over the performance of the partial‐nitritation process and the bacterial community dynamics of the systems analyzed. Ammonium oxidation efficiency decreased with the addition of the amino acid, and a net nitrogen elimination occurred in presence of cysteine through the operation period. Bacterial community dynamics showed a decrease of Nitrosomonas species and a proliferation of putative heterotrophs with nitrification capacity, such as Pseudomonas, or denitrification capacity, such as Denitrobacter or Alicycliphilus. The addition of cysteine irreversible affected the bioreactors, which could not achieve the performance obtained before the addition of the amino acid. A mathematical predictive equation of the process performance depending on cysteine concentration added and operational time under such concentration was developed.

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 ± 0.05 to 6.49 ± 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.

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.