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The high prevalence of antibiotic resistant bacteria (ARB) in several environments is a great concern threatening human health. Particularly, wastewater treatment plants (WWTP) become important contributors to the dissemination of ARB to receiving water bodies, due to the inefficient management or treatment of highly antibiotic-concentrated wastewaters. Hence, it is vital to develop molecular tools that allow proper monitoring of the genes encoding resistances to these important therapeutic compounds (antibiotic resistant genes, ARGs). For an accurate quantification of ARGs, there is a need for sensitive and robust qPCR assays supported by a good design of primers and validated protocols. In this study, eleven relevant ARGs were selected as targets, including aadA and aadB (conferring resistance to aminoglycosides); ampC, blaTEM, blaSHV, and mecA (resistance to beta-lactams); dfrA1 (resistance to trimethoprim); ermB (resistance to macrolides); fosA (resistance to fosfomycin); qnrS (resistance to quinolones); and tetA(A) (resistance to tetracyclines). The in silico design of the new primer sets was performed based on the alignment of all the sequences of the target ARGs (orthology grade > 70%) deposited in the Kyoto Encyclopedia of Genes and Genomes (KEGG) database, allowing higher coverages of the ARGs' biodiversity than those of several primers described to date. The adequate design and performance of the new molecular tools were validated in six samples, retrieved from both natural and engineered environments related to wastewater treatment. The hallmarks of the optimized qPCR assays were high amplification efficiency (> 90%), good linearity of the standard curve (R2 > 0.980), repeatability and reproducibility across experiments, and a wide linear dynamic range. The new primer sets and methodology described here are valuable tools to upgrade the monitorization of the abundance and emergence of the targeted ARGs by qPCR in WWTPs and related environments. [ABSTRACT FROM AUTHOR]

Nowadays, the discharge of wastewater is a global concern due to the damage caused to human and environmental health. Wastewater treatment has progressed to provide environmentally and economically sustainable technologies. The biological treatment of wastewater is one of the fundamental bases of this field, and the employment of new technologies based on granular biofilm systems is demonstrating success in tackling the environmental issues derived from the discharge of wastewater. The granular-conforming microorganisms must be evaluated as functional entities because their activities and functions for removing pollutants are interconnected with the surrounding microbiota. The deep knowledge of microbial communities allows for the improvement in system operation, as the proliferation of microorganisms in charge of metabolic roles could be modified by adjustments to operational conditions. This is why engineering must consider the intrinsic microbiological aspects of biological wastewater treatment systems to obtain the most effective performance. This review provides an extensive view of the microbial ecology of biological wastewater treatment technologies based on granular biofilms for mitigating water pollution. [ABSTRACT FROM AUTHOR]

BACKGROUND: A membrane bioreactor and two hybrid moving bed biofilm reactor‐membrane bioreactor systems were operated in parallel for the treatment of variable‐salinity wastewater. Influent salinity was changed according to tidal‐like cycles: 6 h of maximum salinity (4.5 or 8.5 mS cm−1) followed by 6 h of regular wastewater salinity (around 1 mS cm−1). RESULTS: For the two salinity scenarios, the bioprocesses operated under different conditions of mixed liquor total suspended solids (2500 and 3500 mg L−1), and hydraulic retention time (6, 9.5 and 12 h). The results showed that the bioprocesses had better performance under 4.5 mS cm−1 than under 8.5 mS cm−1 in terms of removal of biological oxygen demand at day 5 (BOD5) (+6.07%), ammonium cation (NH4+) (+41.29%) and total nitrogen (+37.64%), the heterotrophic and autotrophic yield coefficients (+0.02777 mgVSS mgCOD−1 and +0.13990 mgVSS mgN−1), and the autotrophic rate of substrate utilization (+3.39559 mgN L−1 h−1). CONCLUSIONS: The results obtained would become of help for the modeling of membrane‐based systems treating variable saline wastewater. © 2019 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

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 BOD5 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 BOD5 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. [ABSTRACT FROM AUTHOR]

The discovering of anaerobic ammonium oxidation (anammox) process led to the development of autotrophic nitrogen removal systems for the treatment of effluents with low C:N rate. The anammox processes provide an efficient way to remove high concentrations of ammonium compounds from industrial and urban wastewater and covert them to dinitrogen. Nevertheless, recently obtained results suggest new ways for research on autotrophic nitrogen removal system including possibility for low temperature operation, adaptation to high organic matter loads and antibiotics inhibition effect. For these reasons, the prevalence and spatial distribution of anammox communities in autotrophic nitrogen removal wastewater treatment technologies, as well as their role in formation of fixed biofilms, are reviewed here in order to illustrate the present and future significance of these microorganisms in environmental biotechnology. [ABSTRACT FROM AUTHOR]

Aerobic Granular Biomass (AGB) technology is widely used for urban and industrial wastewater treatment, however, its application in groundwater remediation, is practically unknown. A mixture of carbendazim, simazine, and diuron were amended to the nitrate-polluted synthetic groundwater at increasing concentrations to validate the ability of technology to remove both kind of pollutants, pesticides and nitrate which are commonly found in the water resources. The nitrate removal was a success with values below 0.010 g·L-1. The increased concentration of pesticides in the influent did not distort the pattern observed for pesticide removal. Carbendazim was almost completely eliminated, followed by simazine elimination, while diuron showed adsorption-desorption patterns during experimentation. The addition of pesticides had a drastic effect on the basal community conducted by proliferation of Hyphomicrobium and Dokdonella. The pesticide compounds had a negative effect on number of copies for fungal population, while archaeal population was unharmed, according to qPCR results. Denitrifying bacteria need 70 days as acclimatization period for achieving activity values as initial inoculum. The results obtained have shown for the first time the capacity of AGB system to treat groundwater polluted with nitrate and pesticide using low carbon load. Therefore, the results suggested the potential application of AGB technology for the purification of groundwater polluted with both nitrates and pesticides. [Display omitted] • AGB is an efficient technology to treat nitrate and pesticides polluted-groundwater. • Carbendazim and simazine were successfully removed even at high concentrations. • Granular biomass revealed bioadsorption-desorption process with diuron. • Fungal and archaeal populations interchanged roles in presence of pesticides. • Hyphomicrobium was the most dominant phylotype in granules. [ABSTRACT FROM AUTHOR]

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 [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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 Biotechnol. Prog., 31:186-193, 2015 [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Biofouling significantly reduces the performance efficiency of membrane bioreactors due to the colonization and accumulation of a matrix of microorganisms. In order to investigate the effect of ultrasonic frequency on biofilm formation and bacterial ecology at different frequencies, four microfiltration membrane modules were subjected at 0, 20, 30 and 40 kHz in pilot-scales. The results of massive parallel sequencing showed that the application of ultrasonic frequency significantly affected the bacterial population in biofouling and modifies the species richness even at low frequency. This research reveals that the application of ultrasonic frequencies avoids the colonization of Acinetobacter genus, which is considered an initial colonizer of biofouling and clogging in the submerged membrane reactor, fact that caused problems in full-scale wastewater treatment plant. The promotion of Gordonia genus was observed during the application of any ultrasounds frequencies (20,30 and 40 Hz). The compositional statistics data exposed the similarity of bacterial communities under medium-high ultrasonic frequency, while those great distances were found in both low and no-ultrasounds frequencies apply. These results suggest that the application of ultrasounds in membrane bioreactor for treating wastewater could be effective in the potential reduction of biofouling formation in order to improve the to improve the useful life of this system with short periods of application of ultrasound. • Biofouling on membrane treatment using ultrasonic frequency. • The ultrasonic applied to membrane affects to microbial population in biofilm. • Acinetobacter genus was the dominant phylotype in the biofouling. • Gordonia genus showed high resistance against high ultrasonic frequency. [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]

Highlights • Two microbial fuel cells were started-up with cold-adapted inoculum. • COD removal and voltage were similar at different operational temperatures. • Fungi disappeared from the microbial fuel cell at high operational temperature. • Methanogenic archaea proliferated under both temperature conditions. • Blastocatella was the strongest-related phylotype to voltage generation. Abstract Two microbial fuel cells were inoculated with activated sludge from Finland and operated under moderate (25 °C) and low (8 °C) temperatures. Operation under real urban wastewater showed similarities in chemical oxygen demand removal and voltage generated, although moderate temperature supported higher ammonium oxidation. Fungi disappeared in the microbial fuel cell operated at temperature of 25 °C. Archaea domain was dominated by methanogenic archaea at both temperature scenarios. Important differences were observed in bacterial communities between both temperatures, however generating similar voltage. The results supported that the implementation of microbial fuel cells in Nordic countries operating under real conditions could be successful, as well as suggested the flexibility of cold-adapted inoculum for starting-up microbial fuel cells, regardless of the operating temperature of the system, obtaining higher COD removal and voltage generation performances at low temperature than at moderate temperature. [ABSTRACT FROM AUTHOR]

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 , Arcobacte r and Acinetobacte r ( 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. [ABSTRACT FROM AUTHOR]

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 5–3 min 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. [ABSTRACT FROM AUTHOR]

The communities of Bacteria , Archaea and Fungi in a full-scale moving bed biofilm reactor in Rukatunturi (Ruka, Finland, in the Polar Arctic Circle) were analyzed in order to observe their ecological role in the operation of this system at low temperature. The bioreactor achieved efficient removal performances organic matter (>96%) and for ammonium (>95%), however a modest removal for total nitrogen (>63–75%). Bacteria dominant phylotypes changed in relation to the different functions: Trichococcus and Polaromonas for denitrification, Trichococcus and Simplicispira for organic matter removal, and Devosia and Thermomonas for nitrification. Bacteria predicted metagenome related to nitrification confirmed ammonium oxidation in nitrification chamber, and reduction of nitrate and nitrite in the denitrification and organic matter removal chambers. Dynamics of Fungi , dominated by Trichosporonaceae and Ascomycota clones, were linked to aeration conditions but not to substrate concentrations. Archaea communities, dominated by Methanobrevibacter genus and Thermoplasmatales -related clones, were not affected by aeration or substrate concentrations. The results obtained offer a valuable insight into the ecological role of microbial communities in full-scale moving bed biofilm reactor operating under extremely low temperatures. • Bacteria communities were sensitive to aeration and substrate concentrations. • Fungi communities were sensitive to aeration conditions. • Archaea communities were not affected by aeration and substrate concentrations. • NH 4 + and NO 3 − removal occurred in the nitrification and denitrification chambers. • Top contribution of nosZ and nirK occurred in the organic matter removal chamber. [ABSTRACT FROM AUTHOR]