Quorum Quenching in Membrane Bioreactors for Fouling Retardation: Complexity Provides Opportunities.

The occurrence of biofouling restricts the widespread application of membrane bioreactors (MBRs) in wastewater treatment. Regulation of quorum sensing (QS) is a promising approach to control biofouling in MBRs, yet the underlying mechanisms are complex and remain to be illustrated. A fundamental understanding of the relationship between QS and membrane biofouling in MBRs is lacking, which hampers the development and application of quorum quenching (QQ) techniques in MBRs (QQMBRs). While many QQ microorganisms have been isolated thus far, critical criteria for selecting desirable QQ microorganisms are still missing. Furthermore, there are inconsistent results regarding the QQ lifecycle and the effects of QQ on the physicochemical characteristics and microbial communities of the mixed liquor and biofouling assemblages in QQMBRs, which might result in unreliable and inefficient QQ applications. This review aims to comprehensively summarize timely QQ research and highlight the important yet often ignored perspectives of QQ for biofouling control in MBRs. We consider what this "information" can and cannot tell us and explore its values in addressing specific and important questions in QQMBRs. Herein, we first examine current analytical methods of QS signals and discuss the critical roles of QS in fouling-forming microorganisms in MBRs, which are the cornerstones for the development of QQ technologies. To achieve targeting QQ strategies in MBRs, we propose the substrate specificity and degradation capability of isolated QQ microorganisms and the surface area and pore structures of QQ media as the critical criteria to select desirable functional microbes and media, respectively. To validate the biofouling retardation efficiency, we further specify the QQ effects on the physicochemical properties, microbial community composition, and succession of mixed liquor and biofouling assemblages in MBRs. Finally, we provide scale-up considerations of QQMBRs in terms of the debated QQ lifecycle, practical synergistic strategies, and the potential cost savings of MBRs. This review presents the limitations of classic QS/QQ hypotheses in MBRs, advances the understanding of the role of QS/QQ in biofouling development/retardation in MBRs, and builds a bridge between the fundamental understandings and practical applications of QQ technology.

Corrigendum: Effect of oxic and anoxic conditions on intracellular storage of polyhydroxyalkanoate and polyphosphate in Magnetospirillum magneticum strain AMB-1.

[This corrects the article DOI: 10.3389/fmicb.2023.1203805.].

Effect of oxic and anoxic conditions on intracellular storage of polyhydroxyalkanoate and polyphosphate in Magnetospirillum magneticum strain AMB-1.

Magnetotactic bacteria (MTB) are microorganisms widely inhabiting the oxic-anoxic interface of aquatic environments. Beside biomineralizing magnetic nanocrystals, MTBs are able to sequester various chemical elements (e.g., carbon and phosphorus) for the biogenesis of intracellular granules, like polyhydroxyalkanoate (PHA) and polyphosphate (polyP), making them potentially important in biogeochemical cycling. Yet, the environmental controls of intracellular storage of carbon and phosphorus in MTB remain poorly understood. Here, we investigated the influence of oxic, anoxic and transient oxic-anoxic conditions on intracellular storage of PHA and polyP in Magnetospirillum magneticum strain AMB-1. In the incubations with oxygen, transmission electron microscopy revealed intercellular granules highly rich in carbon and phosphorus, which were further interpreted as PHA and polyP based on chemical and Energy-Dispersive X-ray spectroscopy analysis. Oxygen had a strong effect on PHA and polyP storage in AMB-1 cells, as PHA and polyP granules accounted for up to 47 ± 23% and 5.1 ± 1.7% of the cytoplasmic space, respectively, during continuous oxic conditions, while granules disappeared in anoxic incubations. Poly 3-hydroxybutyrate (PHB) and poly 3-hydroxyvalerate (PHV) accounted for 0.59 ± 0.66% and 0.0033 ± 0.0088% of dry cell weight, respectively, in anoxic incubations, while the values increased by a factor of 7 and 37 after oxygen was introduced. The results highlight a tight link between oxygen, carbon and phosphorus metabolisms in MTB, where favorable oxic growth conditions can lead to metabolic induction of polyP and PHA granule biogenesis.

Aerobic biodegradation of quinoline under denitrifying conditions in membrane-aerated biofilm reactor.

Aerobic denitrification is being investigated as a novel biological nitrogen removal process, yet the knowledge on aerobic denitrification is limited to pure culture isolations and its occurrence in bioreactors remains unclear. This study investigated the feasibility and capacity of applying aerobic denitrification in membrane aerated biofilm reactor (MABR) for biological treatment of quinoline-laden wastewater. Stable and efficient removals of quinoline (91.5 ± 5.2%) and nitrate (NO3-) (86.5 ± 9.3%) were obtained under different operational conditions. Enhanced formation and function of extracellular polymeric substances (EPS) were observed at increasing quinoline loadings. MABR biofilm was highly enriched with aerobic quinoline-degrading bacteria, with a predominance of Rhodococcus (26.9 ± 3.7%) and secondary abundance of Pseudomonas (1.7 ± 1.2%) and Comamonas (0.94 ± 0.9%). Metagenomic analysis indicated that Rhodococcus contributed significantly to both aromatic degradation (24.5 ± 21.3%) and NO3- reduction (4.5 ± 3.9%), indicating its key role in aerobic denitrifying quinoline biodegradation. At increasing quinoline loadings, abundances of aerobic quinoline degradation gene oxoO and denitrifying genes of napA, nirS and nirK increased; there was a significant positive correlation of oxoO with nirS and nirK (p < 0.05). Aerobic quinoline degradation was likely initiated by hydroxylation, encoded by oxoO, followed by stepwise oxidations through 5,6-dihydroxy-1H-2-oxoquinoline or 8-hydroxycoumarin pathway. The results advance our understanding of quinoline degradation during biological nitrogen removal, and highlight the potential implementation of aerobic denitrification driven quinoline biodegradation in MABR for simultaneous removal of nitrogen and recalcitrant organic carbon from coking, coal gasification and pharmaceutical wastewaters.

Abiotic transformations of sulfamethoxazole by hydroxylamine, nitrite and nitric oxide during wastewater treatment: Kinetics, mechanisms and pH effects.

Hydroxylamine (NH2OH), nitrite (NO2-) and nitric oxide (NO), intermediates enzymatically formed during biological nitrogen removal processes, can engage in chemical (abiotic) transformations of antibiotics. This study determined the kinetics, mechanisms and pathways of abiotic transformations of the antibiotic sulfamethoxazole (SMX) by NH2OH, NO2- and NO in a series of batch tests under different pH and oxygen conditions. While NH2OH was not able to directly transform SMX, NO2- (with HNO2 as the actual reactant) and NO can chemically transform SMX primarily through hydroxylation, nitration, deamination, nitrosation, cleavage of S-N, N-C and C-S bonds, and coupling reactions. There were substantial overlaps in transformation product formations during abiotic transformations by HNO2- and NO. The second order rate constants of SMX with NO2- and NO were determined in the range of 1.5 × 10-1 - 4.8 × 103 M-1 s-1 and 1.0 × 102 - 3.1 × 104 M-1 s-1, respectively, under varying pH (4 - 9) and anoxic or oxic conditions. Acidic pH significantly enhanced abiotic transformation kinetics, and facilitated nitration, nitrosation, and cleavage of S-N and N-C bonds. The findings advance our understanding of the fate of antibiotics during biological nitrogen removal, and highlight the role of enzymatically formed reactive nitrogen species in the antibiotic degradation.

Development and characterizations of hydrogenotrophic denitrification granular process: Nitrogen removal capacity and adaptability.

Hydrogenotrophic denitrification (HD) is a promising autotrophic biological process for advanced nitrogen removal, while sludge granulation was seldom reported. This study aimed to cultivate granular sludge to improve capacity and stability of HD process. The resulting HD granular sludge performed high nitrogen removal rate (NRR) of 0.42 ± 0.0.4 kgN/(m3·d) with low accumulation of nitrite and nitrous oxide emission. HD granular sludge reactor performed over 3 times higher NRR compared to that in HD fixed-bed biofilm reactor (0.13 ± 0.01 kgN/(m3·d). Besides, granular sludge reactor could treat groundwater well even at the low temperature of 15 °C. The dominant genera were Hydrogenophaga and Comamonas in granular sludge, and Dechloromonas in biofilm. Noticeably, sulfate in the groundwater stimulated the growth of sulfur converting microbes with increasing abundances of sulfite reductase gene and sulfate-reducing bacteria Desulfovibrio. This study highlights the potential implementation of HD process in granular sludge reactor for advance nitrogen removal from impaired groundwater.

Microbial bioremediation of produced water under different redox conditions in marine sediments.

The discharge of produced water from offshore oil platforms is an emerging concern due to its potential adverse effects on marine ecosystems. In this study, we investigated the feasibility and capability of using marine sediments for the bioremediation of produced water. We utilized a combination of porewater and solid phase analysis in a series of sediment batch incubations amended with produced water and synthetic produced water to determine the biodegradation of hydrocarbons under different redox conditions. Significant removal of benzene, toluene, ethylbenzene and xylene (BTEX) compounds was observed under different redox conditions, with biodegradation efficiencies of 93-97% in oxic incubations and 45-93% in anoxic incubations with nitrate, iron oxide or sulfate as the electron acceptor. Higher biodegradation rates of BTEX were obtained by incubations dominated by nitrate reduction (104-149 nmolC/cm3/d) and oxygen respiration (52-57 nmolC/cm3/d), followed by sulfate reduction (14-76 nmolC/cm3/d) and iron reduction (29-39 nmolC/cm3/d). Chemical fingerprint analysis showed that hydrocarbons were biodegraded to smaller alcohols/acids under oxic conditions compared to anoxic conditions with nitrate, indicating that the presence of oxygen facilitated a more complete biodegradation process. Toxicity of treated produced water to the marine copepod Acartia tonsa was reduced by half after sediment incubations with oxygen and nitrate. Our study emphasizes the possibility to use marine sediment as a biofilter for treating produced water at sea without extending the oil and gas platform or implementing a large-scale construction.

Exploring the role of heterotrophs in partial nitritation-anammox process treating thermal hydrolysis process - anaerobic digestion reject water.

Heterotrophic bacteria (HB) are generally prevalent in anammox-based processes, but their functional and ecological roles in partial nitritation-anammox (PN/A) process treating high-organics wastewater remained unclear. This study aimed to elucidate HB activities and microbial interactions in a one-stage PN/A treating thermal hydrolysis process (THP) - anaerobic digestion (AD) reject water. The PN/A reactor achieved a satisfactory nitrogen removal rate of 0.58 ± 0.06 g N/(L·d), and around 12% of COD in the THP-AD reject water was removed. N2O emission factors of the PN/A reactor were 1.15% ± 0.18% treating synthetic wastewater, and 0.95% ± 0.06% treating reject water. A balanced symbiotic relationship was maintained between HB and functional groups (i.e., anammox bacteria and aerobic-ammonia-oxidizing bacteria) over the reactor operation. The relative abundances of Anaerolineae spp. clearly increased, while Denitratisoma, capable of denitrification, slightly decreased when treating THP-AD reject water. The preference for electron donors of heterotrophs explained discrepant growth trends.

Role of Ammonia Oxidation in Organic Micropollutant Transformation during Wastewater Treatment: Insights from Molecular, Cellular, and Community Level Observations.

Organic micropollutants (OMPs) are a threat to aquatic environments, and wastewater treatment plants may act as a source or a barrier of OMPs entering the environment. Understanding the fate of OMPs in wastewater treatment processes is needed to establish efficient OMP removal strategies. Enhanced OMP biotransformation has been documented during biological nitrogen removal and has been attributed to the cometabolic activity of ammonia-oxidizing bacteria (AOB) and, specifically, to the ammonia monooxygenase (AMO) enzyme. Yet, the exact mechanisms of OMP biotransformation are often unknown. This critical review aims to fundamentally and quantitatively evaluate the role of ammonia oxidation in OMP biotransformation during wastewater treatment processes. OMPs can be transformed by AOB via direct and indirect enzymatic reactions: AMO directly transforms OMPs primarily via hydroxylation, while biologically produced reactive nitrogen species (hydroxylamine (NH2OH), nitrite (NO2-), and nitric oxide (NO)) can chemically transform OMPs through nitration, hydroxylation, and deamination and can contribute significantly to the observed OMP transformations. OMPs containing alkyl, aliphatic hydroxyl, ether, and sulfide functional groups as well as substituted aromatic rings and aromatic primary amines can be biotransformed by AMO, while OMPs containing alkyl groups, phenols, secondary amines, and aromatic primary amines can undergo abiotic transformations mediated by reactive nitrogen species. Higher OMP biotransformation efficiencies and rates are obtained in AOB-dominant microbial communities, especially in autotrophic reactors performing nitrification or nitritation, than in non-AOB-dominant microbial communities. The biotransformations of OMPs in wastewater treatment systems can often be linked to ammonium (NH4+) removal following two central lines of evidence: (i) Similar transformation products (i.e., hydroxylated, nitrated, and desaminated TPs) are detected in wastewater treatment systems as in AOB pure cultures. (ii) Consistency in OMP biotransformation (rbio, µmol/g VSS/d) to NH4+ removal (rNH4+, mol/g VSS/d) rate ratios (rbio/rNH4+) is observed for individual OMPs across different systems with similar rNH4+ and AOB abundances. In this review, we conclude that AOB are the main drivers of OMP biotransformation during wastewater treatment processes. The importance of biologically driven abiotic OMP transformation is quantitatively assessed, and functional groups susceptible to transformations by AMO and reactive nitrogen species are systematically classified. This critical review will improve the prediction of OMP transformation and facilitate the design of efficient OMP removal strategies during wastewater treatment.

Abiotic Nitrous Oxide (N2O) Production Is Strongly pH Dependent, but Contributes Little to Overall N2O Emissions in Biological Nitrogen Removal Systems.

Hydroxylamine (NH2OH) and nitrite (NO2-), intermediates during the nitritation process, can engage in chemical (abiotic) reactions that lead to nitrous oxide (N2O) generation. Here, we quantify the kinetics and stoichiometry of the relevant abiotic reactions in a series of batch tests under different and relevant conditions, including pH, absence/presence of oxygen, and reactant concentrations. The highest N2O production rates were measured from NH2OH reaction with HNO2, followed by HNO2 reduction by Fe2+, NH2OH oxidation by Fe3+, and finally NH2OH disproportionation plus oxidation by O2. Compared to other examined factors, pH had the strongest effect on N2O formation rates. Acidic pH enhanced N2O production from the reaction of NH2OH with HNO2 indicating that HNO2 instead of NO2- was the reactant. In departure from previous studies, we estimate that abiotic N2O production contributes little (< 3% of total N2O production) to total N2O emissions in typical nitritation reactor systems between pH 6.5 and 8. Abiotic contributions would only become important at acidic pH (≤ 5). In consideration of pH effects on both abiotic and biotic N2O production pathways, circumneutral pH set-points are suggested to minimize overall N2O emissions from nitritation systems.

The effect of pH on N2O production in intermittently-fed nitritation reactors.

The effect of pH on nitrous oxide (N2O) production rates was quantified in an intermittently-fed lab-scale sequencing batch reactor performing high-rate nitritation. N2O and other nitrogen (N) species (e.g. ammonium (NH4+), nitrite, hydroxylamine and nitric oxide) were monitored to identify in-cycle dynamics and determine N conversion rates at controlled pH set-points (6.5, 7, 7.5, 8 and 8.5). Operational conditions and microbial compositions remained similar during long-term reactor-scale pH campaigns. The specific ammonium removal rates and nitrite accumulation rates varied little with varying pH levels (p > 0.05). The specific net N2O production rates and net N2O yield of NH4+ removed (ΔN2O/ΔNH4+) increased up to seven-fold from pH 6.5 to 8, and decreased slightly with further pH increase to 8.5 (p < 0.05). Best-fit model simulations predicted nitrifier denitrification as the dominant N2O production pathway (≥87% of total net N2O production) at all examined pH. Our study highlights the effect of pH on biologically mediated N2O emissions in nitrogen removal systems and its importance in the design of N2O mitigation strategies.

Adaption and recovery of Nitrosomonas europaea to chronic TiO2 nanoparticle exposure.

Although the adverse impacts of emerging nanoparticles (NPs) on the biological nitrogen removal (BNR) process have been broadly reported, the adaptive responses of NP-impaired nitrifiers and the related mechanisms have seldom been addressed to date. Here, we systematically explored the adaption and recovery capacities of the ammonia oxidizer Nitrosomonas europaea under chronic TiO2 NP exposure and different dissolved oxygen (DO) conditions at the physiological and transcriptional levels in a chemostat reactor. N. europaea cells adapted to 50 mg/L TiO2 NP exposure after 40-d incubation and the inhibited cell growth, membrane integrity, nitritation rate, and ammonia monooxygenase activity all recovered regardless of the DO concentrations. Transmission electron microscope imaging indicated the remission of the membrane distortion after the cells' 40-d adaption to the NP exposure. The microarray results further suggested that the metabolic processes associated with the membrane repair were pivotal for cellular adaption/recovery, such as the membrane efflux for toxicant exclusion, the structural preservation or stabilization, and the osmotic equilibrium adjustment. In addition, diverse metabolic and stress-defense pathways, including aminoacyl-tRNA biosynthesis, respiratory chain, ATP production, toxin-antitoxin 'stress-fighting', and DNA repair were activated for the cellular adaption coupled with the metabolic activity recovery, probably via recovering the energy production/conversion efficiency and mediating the non-photooxidative stress. Finally, low DO (0.5 mg/L) incubated cells were more susceptible to TiO2 NP exposure and required more time to adapt to and recover from the stress, which was probably due to the stimulation limitation of the oxygen-dependent energy metabolism with a lower oxygen supply. The findings of this study provide new insights into NP contamination control and management adjustments during the BNR process.

The pH dependency of N-converting enzymatic processes, pathways and microbes: effect on net N2 O production.

Nitrous oxide (N2 O) is emitted during microbiological nitrogen (N) conversion processes, when N2 O production exceeds N2 O consumption. The magnitude of N2 O production vs. consumption varies with pH and controlling net N2 O production might be feasible by choice of system pH. This article reviews how pH affects enzymes, pathways and microorganisms that are involved in N-conversions in water engineering applications. At a molecular level, pH affects activity of cofactors and structural elements of relevant enzymes by protonation or deprotonation of amino acid residues or solvent ligands, thus causing steric changes in catalytic sites or proton/electron transfer routes that alter the enzymes' overall activity. Augmenting molecular information with, e.g., nitritation or denitrification rates yields explanations of changes in net N2 O production with pH. Ammonia oxidizing bacteria are of highest relevance for N2 O production, while heterotrophic denitrifiers are relevant for N2 O consumption at pH > 7.5. Net N2 O production in N-cycling water engineering systems is predicted to display a 'bell-shaped' curve in the range of pH 6.0-9.0 with a maximum at pH 7.0-7.5. Net N2 O production at acidic pH is dominated by N2 O production, whereas N2 O consumption can outweigh production at alkaline pH. Thus, pH 8.0 may be a favourable pH set-point for water treatment applications regarding net N2 O production.

Low nitrous oxide production through nitrifier-denitrification in intermittent-feed high-rate nitritation reactors.

Nitrous oxide (N2O) production from autotrophic nitrogen conversion processes, especially nitritation systems, can be significant, requires understanding and calls for mitigation. In this study, the rates and pathways of N2O production were quantified in two lab-scale sequencing batch reactors operated with intermittent feeding and demonstrating long-term and high-rate nitritation. The resulting reactor biomass was highly enriched in ammonia-oxidizing bacteria, and converted ∼93 ± 14% of the oxidized ammonium to nitrite. The low DO set-point combined with intermittent feeding was sufficient to maintain high nitritation efficiency and high nitritation rates at 20-26 °C over a period of ∼300 days. Even at the high nitritation efficiencies, net N2O production was low (∼2% of the oxidized ammonium). Net N2O production rates transiently increased with a rise in pH after each feeding, suggesting a potential effect of pH on N2O production. In situ application of 15N labeled substrates revealed nitrifier denitrification as the dominant pathway of N2O production. Our study highlights operational conditions that minimize N2O emission from two-stage autotrophic nitrogen removal systems.