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The present work aims to use aerobic granular sludge technology for the treatment of wastewater containing high organic matter loads and a mixture of phenolic compounds normally present in olive washing water. The physicochemical performance of five bioreactors treating different concentrations of mixture of phenolic acid was monitored to observe the response of the systems. The bioreactors that operated at 50, 100 and 300 mg L -1 did not show relevant changes in terms of performance and granules properties, showing high ratio of phenolic compound removal ratio. However, the bioreactors operated with high phenolic compound concentrations showed low rates of organic matter, nitrogen and phenolic acid removal. In the same way, high concentrations of phenolic compounds determined the disintegration of the granular biomass. Next-generation sequencing studies showed a stable community structure in the bioreactors operating with 50, 100 and 300 mg L -1 of phenolic acids, with the genera Lampropedia and Arenimonas, family Xanthobacteraceae and Fungi Pezizomycotina as the dominant phylotypes. Conversely, the reactors operated at 500 and 600 mg L -1 of phenolic substances promoted the proliferation of Oligohymenophorea ciliates. Thus, this study suggests that aerobic granular sludge technology could be useful for the treatment of wastewaters such as olive washing water., (Copyright © 2019 Elsevier B.V. All rights reserved.)

A lab-scale partial nitritation SBR was operated at 11 °C for 300 days used for the treatment of high-ammonium wastewater, which was inoculated with activated sludge from Rovaniemi WWTP (located in Polar Arctic Circle) in order to evaluate the influence the temperature on the performance, stability and dynamics of its microbial community. The partial nitritation achieved steady-state long-term operation and granulation process was not affected despite the low temperature and high ammonia concentration. The steady conditions were reached after 60 days of operation where the granular biomass was fully-formed and the 50%-50% of ammonium-nitrite effluent was successful achieved. Inoculation with cold adapted inoculum showed to yield bigger, denser granules with faster start-up without necessity of low temperature adaptation period. Next-generation sequences techniques showed that Trichosporonaceae and Xanthomonadaceae were the dominant OTUs in the mature granules. Our study could be useful in the implementation of full-scale partial nitritation reactors in cold regions such as Nordic countries for treating wastewater with high concentration of ammonium., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

A membrane bioreactor and two hybrid moving bed bioreactor-membrane bioreactors were operated for the treatment of variable salinity wastewater, changing in cycles of 6-h wastewater base salinity and 6-h maximum salinity (4.5 and 8.5 mS cm -1 electric conductivity, which relate to 2.4 and 4.8 g L -1 NaCl, respectively), under different hydraulic retention times (6, 9.5, and 12 h) and total solids concentrations (2500 and 3500 mg L -1 ). The evaluation of the performance of the systems showed that COD removal performance was unaffected by salinity conditions, while BOD 5 and TN removals were significantly higher in the low-salinity scenario. The microbial community structure showed differences with respect to salinity conditions for Eukarya, suggesting their higher sensitivity for salinity with respect to Prokarya, which were similar at both salinity scenarios. Nevertheless, the intra-OTU distribution of consistently represented OTUs of Eukarya and Prokarya was affected by the different salinity maximums. Multivariate redundancy analyses showed that several genera such as Amphiplicatus (0.01-5.90%), Parvibaculum (0.27-1.19%), Thiothrix (0.30-1.19%), Rhodanobacter (2.81-5.85%), Blastocatella (0.21-2.01%), and Nitrobacter (0.80-0.99%) were positively correlated with BOD 5 and TN removal, and the ecological roles of these were proposed. All these genera were substantially more represented under low-salinity conditions (10-500% higher relative abundance), demonstrating that they might be of importance for the treatment of variable salinity wastewater. Evaluation of Eukarya OTUs showed that many of them lack a consistent taxonomic classification, which highlights the lack of knowledge of the diversity and ecological role of Eukaryotes in saline wastewater treatment processes. The results obtained will be of interest for future design and operation of salinity wastewater treatment systems particularly because little is known on the effect of variable salinity conditions in wastewater treatment.

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., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

The nitrogen cycle has been expanded with the recent discovery of Nitrospira strains that can conduct complete ammonium oxidation (commamox). Their importance in the nitrogen cycle within engineered ecosystems has not yet been analyzed. In this research, the community structure of the Bacteria domain of six full-scale activated sludge systems and three autotrophic nitrogen removal systems in the Netherlands and China has been investigated through tag-454-pyrosequencing. The phylogenetic analyses conducted in the present study showed that just a few of the Nitrospira sequences found in the bioreactors were comammox. Multivariate redundancy analysis of nitrifying genera showed an outcompetition of Nitrosomonas and non-comammox Nitrospira. Operational data from the bioreactors suggested that comammox could be favored at low temperature, low nitrogen substrate, and high dissolved oxygen. The non-ubiquity and low relative abundance of comammox in full-scale bioreactors suggested that this phylotype is not very relevant in the nitrogen cycle in wastewater treatment plants.

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. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1254-1263, 2016., (© 2016 American Institute of Chemical Engineers.)

Two-stage technologies have been developed for anaerobic digestion of waste-activated sludge. In this study, the archaeal and bacterial community structure dynamics and bioprocess performance of a bench-scale two-stage anaerobic digester treating urban sewage sludge have been studied by the means of high-throughput sequencing techniques and physicochemical parameters such as pH, dried sludge, volatile dried sludge, acid concentration, alkalinity, and biogas generation. The coupled analyses of archaeal and bacterial communities and physicochemical parameters showed a direct relationship between archaeal and bacterial populations and bioprocess performance during start-up and working operation of a two-stage anaerobic digester. Moreover, results demonstrated that archaeal and bacterial community structure was affected by changes in the acid/alkalinity ratio in the bioprocess. Thus, a predominance of the acetoclastic methanogen Methanosaeta was observed in the methanogenic bioreactor at high-value acid/alkaline ratio, while a predominance of Methanomassilicoccaeceae archaea and Methanoculleus genus was observed in the methanogenic bioreactor at low-value acid/alkaline ratio. Biodiversity tag-iTag sequencing studies showed that methanogenic archaea can be also detected in the acidogenic bioreactor, although its biological activity was decreased after 4 months of operation as supported by physicochemical analyses. Also, studies of the VFA producers and VFA consumers microbial populations showed as these microbiota were directly affected by the physicochemical parameters generated in the bioreactors. We suggest that the results obtained in our study could be useful for future implementations of two-stage anaerobic digestion processes at both bench- and full-scale.

It has been demonstrated that an anaerobic digestion process cannot attain an efficient removal of several amino acids, with methionine being one of the most persistent of these. Thus, the effect that methionine amino acid has over the partial-nitritation process with fixed-biofilm configuration in terms of performance and bacterial community dynamics has been investigated. With respect to the performance with no addition, 100 mg/L methionine loading decreased ammonium oxidation efficiency in 60% and 100% at concentrations of 300 and 500 mg/L methionine, respectively. Bacterial biomass sharply increased by 30, 65, and 230% with the addition of 100, 300, and 500 mg/L methionine, respectively. Bacterial community analysis showed that methionine addition supported the proliferation of a diversity of heterotrophic genera, such as Lysobacter and Micavibrio, and reduced the relative abundance of ammonium oxidizing genus Nitrosomonas. This research shows that the addition of methionine affects the performance of the partial-nitritation process. In this sense, amino acids can pose a threat for the of partial-nitritation process treating anaerobic digester supernatant at full-scale implementation.

Bacterial community structure of full-scale anammox bioreactor is still mainly unknown. It has never been analyzed whether different anammox bioreactor configurations might result in the development of different bacterial community structures among these systems. In this work, the bacterial community structure of six full-scale autotrophic nitrogen removal bioreactors located in The Netherlands and China operating under three different technologies and with different influent wastewater characteristics was studied by the means of pyrotag sequencing evaluation of the bacterial assemblage yielded a great diversity in all systems. The most represented phyla were the Bacteroidetes and the Proteobacteria, followed by the Planctomycetes. 14 OTUs were shared by all bioreactors, but none of them belonged to the Brocadiales order. Statistical analysis at OTU level showed that differences in the microbial communities were high, and that the main driver of the bacterial assemblage composition was different for the distinct phyla identified in the six bioreactors, depending on bioreactor technology or influent wastewater characteristics., (© 2015 American Institute of Chemical Engineers.)

Full-scale applications of autotrophic nitrogen removal technologies for the treatment of digested sludge liquor have proliferated during the last decade. Among these technologies, the aerobic/anoxic deammonification process (DEMON) is one of the major applied processes. This technology achieves nitrogen removal from wastewater through anammox metabolism inside a single bioreactor due to alternating cycles of aeration. To date, microbial community composition of full-scale DEMON bioreactors have never been reported. In this study, bacterial community structure of a full-scale DEMON bioreactor located at the Apeldoorn wastewater treatment plant was analyzed using pyrosequencing. This technique provided a higher-resolution study of the bacterial assemblage of the system compared to other techniques used in lab-scale DEMON bioreactors. Results showed that the DEMON bioreactor was a complex ecosystem where ammonium oxidizing bacteria, anammox bacteria and many other bacterial phylotypes coexist. The potential ecological role of all phylotypes found was discussed. Thus, metagenomic analysis through pyrosequencing offered new perspectives over the functioning of the DEMON bioreactor by exhaustive identification of microorganisms, which play a key role in the performance of bioreactors. In this way, pyrosequencing has been proven as a helpful tool for the in-depth investigation of the functioning of bioreactors at microbiological scale.

Autotrophic nitrogen removal technologies have proliferated through the last decade. Among these, a promising one is the membrane bioreactor (MBR) Anammox, which can achieve very high solids retention time and therefore sets a proper environment for the cultivation of anammox bacteria. In this sense, the MBR Anammox is an efficient technology for the treatment of effluents with low organic carbon and high ammonium concentrations once it has been treated under partial nitrification systems. A lab-scale MBR Anammox bioreactor has been built at the Technological University of Delft, The Netherlands and has been proven for efficient nitrogen removal and efficient cultivation of anammox bacteria. In this study, next-generation sequencing techniques have been used for the investigation of the bacterial communities of this MBR Anammox for the first time ever. A strong domination of Candidatus Brocadia bacterium and also the presence of a myriad of other microorganisms that have adapted to this environment were detected, suggesting that the MBR Anammox bioreactor might have a more complex microbial ecosystem that it has been thought. Among these, nitrate-reducing heterotrophs and primary producers, among others, were identified. Definition of the ecological roles of the OTUs identified through metagenomic analysis was discussed., (© 2014 American Institute of Chemical Engineers.)

Identification of anaerobic ammonium oxidizing (anammox) bacteria by molecular tools aimed at the evaluation of bacterial diversity in autotrophic nitrogen removal systems is limited by the difficulty to design universal primers for the Bacteria domain able to amplify the anammox 16S rRNA genes. A metagenomic analysis (pyrosequencing) of total bacterial diversity including anammox population in five autotrophic nitrogen removal technologies, two bench-scale models (MBR and Low Temperature CANON) and three full-scale bioreactors (anammox, CANON, and DEMON), was successfully carried out by optimization of primer selection and PCR conditions (annealing temperature). The universal primer 530F was identified as the best candidate for total bacteria and anammox bacteria diversity coverage. Salt-adjusted optimum annealing temperature of primer 530F was calculated (47°C) and hence a range of annealing temperatures of 44-49°C was tested. Pyrosequencing data showed that annealing temperature of 45°C yielded the best results in terms of species richness and diversity for all bioreactors analyzed.

The ability of bacteria isolated from a fixed-film bioreactor to precipitate phosphate crystals for the treatment of domestic wastewater in both artificial and natural media was studied. When this was demonstrated in artificial solid media for crystal formation, precipitation took place rapidly, and crystal formation began 3 days after inoculation. The percentage of phosphate-forming bacteria was slightly higher than 75%. Twelve major colonies with phosphate precipitation capacity were the dominant heterotrophic platable bacteria growing aerobically in artificial media. According to their taxonomic affiliations (based on partial sequencing of the 16S rRNA), the 12 strains belonged to the following genera of Gram-negative bacteria: Rhodobacter, Pseudoxanthobacter, Escherichia, Alcaligenes, Roseobacter, Ochrobactrum, Agromyce, Sphingomonas and Paracoccus. The phylogenetic tree shows that most of the identified populations were evolutionarily related to the Alphaproteobacteria (91.66% of sequences). The minerals formed were studied by X-ray diffraction, scanning electron microscopy (SEM), and energy dispersive X-ray microanalysis (EDX). All of these strains formed phosphate crystals and precipitated struvite (MgNH4PO4·6H2O), bobierrite [Mg3(PO4)2·8H2O] and baricite [(MgFe)3(PO4)2·8H2O]. The results obtained in this study show that struvite and spherulite crystals did not show any cell marks. Moreover, phosphate precipitation was observed in the bacterial mass but also near the colonies. Our results suggest that the microbial population contributed to phosphate precipitation by changing the media as a consequence of their metabolic activity. Moreover, the results of this research suggest that bacteria play an active role in the mineral precipitation of soluble phosphate from urban wastewater in submerged fixed-film bioreactors.

In order to evaluate the effect of flux on membrane fouling, the performance of a bench-scale submerged membrane bioreactor (MBR) equipped with ultrafiltration membranes (ZENON) was investigated under real conditions at different flux rates. The pilot plant was located at the wastewater treatment plant of the city of Granada (Spain). Influent used in the experiments came from the primary settling tank. Assays carried out under different operating conditions indicated that dTMP/dt increased in accordance with the increase in flux. The results showed a significant impact on the rate of transmembrane pressure, while the behavior of membrane fouling was logarithmic with respect to the flux. These findings could be of some importance for understanding the behavior of the membrane, since over 20.57 L m(-2) h(-1) the flux rate produced a significant increase in transmembrane pressure. The data therefore suggest that an increase in the net flux significantly affects membrane fouling.

Granular activated sludge has been described as a promising tool in treating wastewater. However, the effect of high concentrations of sulphur amino acids, cysteine and methionine, in the evolution, development and stability of AGS-SBRs (aerobic granular sludge in sequential batch reactors) and their microbial communities is not well-established. Therefore, this study aimed to evaluate microbial communities' size, structure and dynamics in two AGS-SBRs fed with two different concentrations of amino acids (50 and 100 mg L−1 of both amino acids). In addition, the impact of the higher level of amino acids was also determined under an acclimatization or shock strategy. While N removal efficiency decreased with amino acids, the removal of the organic matter was generally satisfactory. Moreover, the abrupt presence of both amino acids reduced even further the removal performance of N, whereas under progressive adaptation, the removal yield was higher. Besides, excellent removal rates of cysteine and methionine elimination were found, in all stages below 80% of the influent values. Generally considered, the addition of amino acids weakly impacts the microbial communities' total abundances. On the contrary, the presence of amino acids sharply modulated the dominant bacterial structures. Furthermore, the highest amino acid concentration under the shock strategy resulted in a severe change in the structure of the microbial community. Acidovorax, Flavobacterium, Methylophilus, Stenotrophomonas and Thauera stood out as the prominent bacteria to cope with the high presence of cysteine and methionine. Hence, the AGS-SBR technology is valuable for treating influents enriched in sulphur Aa inclusively when a shock strategy was used. [ABSTRACT FROM AUTHOR]

A membrane bioreactor and two hybrid moving bed biofilm reactor-membrane bioreactor systems were operated for the treatment of wastewater with tidal salinity fluctuations under hydraulic retention times of 6, 9.5 and 12 h, and operational solids concentrations of around 2500 and 3500 mg L −1 . The three configurations were studied in terms of carbon and nitrogen removal, heterotrophic and autotrophic kinetics, and bacterial community structure. The performance of the systems was good in terms of organic matter removal – between 85–100% and 95–100% for COD and BOD 5 , respectively, showing higher efficiencies at higher solids concentration and hydraulic retention times. Nitrogen removal obtained was in the range of 30–50%. The bacterial community structure of the suspended and attached biomass in the systems was more influenced by the operational solids (80% clustering cutoff) and hydraulic retention time (60% clustering cutoff) than by technological configuration (40% clustering cutoff), as determined by ordination analyses. Massive parallel sequencing showed the presence of Nitrobacter and Rhodanobacter at almost all operation scenarios, and thus were identified as major players in treatment of tidal salinity variation wastewater. Overall, this research proved that the MBR and hybrid MBBR-MBR systems could successfully treat urban wastewater subjected to tidal salinity variations. Nevertheless, more research is needed in order to enhance nitrogen removal performance under these conditions. [ABSTRACT FROM AUTHOR]

An autotrophic nitrogen removal bioreactor with granular biomass was exposed to high antibiotics concentration in order to evaluate its impact over the performance and the biomass of this bioprocess. A mixture of azithromycin, norfloxacin, trimethoprim and sulfamethoxazole caused loss of autotrophic nitrogen removal performance, coupled to a deep change in the bacterial community diversity and structure of the granular biomass. Azithromycin, norfloxacin and trimethoprim were efficiently removed in the CANON bioreactor, reducing its concentration 77.9 ± 11.2%, 51.7 ± 10.7% and 57.8 ± 8.1%, respectively. The granular biomass changed significantly with the addition of the antibiotics, decreasing in settling velocity but increasing in compactness, losing its inner porous structure but developing a protective outer layer build of cell material. Prolonged operation under the antibiotics loading promoted the adaptation of multi-drug resistant fungus Scedosporium boydii fungal species and of Acidovorax ebreus TPSY , Alcaligenes aquatilis , Paracoccus versutus or Ochrobactrum antropii , which have been identified as human, animal and/or plant pathogens. [ABSTRACT FROM AUTHOR]