Nitrogen removal performance of a hybrid anammox reactor

The anaerobic ammonium oxidation (anammox) process has attracted considerable attention in recent years as an alternative to conventional nitrogen removal technologies. In this study, an innovative hybrid reactor combining fluidized and fixed beds for anammox treatment was developed. The fluidized bed was mechanically stirred and the gaseous product could be rapidly released from the anammox sludge to prevent washout of the sludge caused by floatation. The fixed bed comprising a non-woven biomass carrier could efficiently catch sludge to reduce washout. During the operation, nitrogen loading rates to the reactor were increased to 27.3 kg N/m³/d, with total nitrogen removal efficiencies of 75%. The biomass concentration in the fluidized bed reached 26-g VSS/L. Anammox granules were observed in the reactors, with settling velocities and sludge volumetric index of 27.3 ± 6.5 m/h and 23 mL/g, respectively. Quantification of extracellular polymeric substances revealed the anammox granules contained a significant amount of extracellular proteins.     

Advances in aerobic granule formation and granule stability in the course of storage and reactor operation

Aerobic granulation is drawing increasing global interest in a quest for an efficient and innovative technology in wastewater treatment. Developed less than two decades ago, extensive research work on aerobic granulation has been reported. The instability of the granule, which is one of the main problems that hinder practical application of aerobic granulation technology, is still to be resolved. This paper presents a review of the literature in aerobic granulation focusing on factors that influence granule formation, granule development and their stability in the context of sludge granulation. The review attempts to shed light on the potential of developing granules with adequate structural stability for practical applications. The possibilities and perspective of using stored granule as inoculums for rapid startup, and as microbial supplement to enhance treatment of bioreactor systems are also discussed.     

Fast start-up, performance and microbial community in a pilot-scale anammox reactor seeded with exotic mature granules

The possibility to introduce the exotic anammox sludge to seed the pilot-scale anammox granular reactor and its fast start-up for treating high nitrogen concentration wastewater were evaluated in this study. The reactor was started up successfully in two weeks; in addition, high nitrogen removal was achieved for a long period. Stoichiometry molar ratios of nitrite conversion and nitrate production to ammonium conversion were calculated to be 1.26 ± 0.02:1 and 0.26 ± 0.01:1, respectively. The Stover-Kincannon model which was first applied in granular anammox process indicated that the granular anammox reactor possessed high nitrogen removal potential of 27.8 kg/m³/d. The anammox granules in the reactor were characterized via microscope observation and fluorescence in situ hybridization technique. Moreover, the microbial community of the granules was quantified to be composed of 91.4-92.4% anammox bacteria by real-time polymerase chain reaction. This pilot study can elucidate further information for industrial granular anammox application.     

PVA-gel beads enhance granule formation in a UASB reactor

PVA-gel beads were used as a biocarrier in a lab-scale UASB reactor treating synthetic wastewater composed of corn steep liquor (CSL) with the aim of evaluating its use as a growth nucleus to enhance granule formation. Over 117 days of operation, the organic loading rate was increased to 22.5kgCOD/m(3)/day with an influent COD of about 10.8g/L at an HRT of 12h with COD removal efficiencies greater than 87%. By the end of the study period, the PVA-gel turned black and granule formation was achieved as compared with the formation of much fewer natural granules without the PVA-gel nucleus. No filamentous bacteria were found on the surface or interior of the PVA-gel beads. The PVA-gel granules had an average settling velocity 200m/h (5cm/s), and a biomass attachment of 0.93g VSS/g PVA-gel. The required time for formation of PVA-gel granules was thus demonstrated to be shorter than that of ordinary sludge granules under the experimental conditions used in this study.     

Selective stimulation of L-arginine uptake contributes to shear stress-induced formation of nitric oxide

The aim of this study was to investigate the kinetics of L-arginine transport mechanisms and the role of extracellular L-arginine in nitric oxide formation during shear stress activation of endothelial cells. Porcine aortic endothelial cells were grown to confluence and were exposed to various amounts of shear stress for 40 min. Formation of nitric oxide was monitored by measuring elevation of endothelial cGMP. Activity of amino acid transport systems was determined by measuring the uptake of L-[3H]leucine (L system) and L-[3H]arginine (y+) under resting and shear stress condition. Shear stress-mediated nitric oxide formation critically depended on the presence of extracellular L-arginine, which increased shear stress-induced cGMP increases in a concentration dependent manner (EC50=123 microM). In addition, shear stress increased L-arginine uptake, while the transport capacity for neutral amino acids (L system) remained unchanged under shear stress conditions. Analysis of the kinetics of the uptake of L-arginine under resting and shear stress conditions indicate that shear stress increased velocity of the high affinity, low capacity transport (y+) without affecting affinity of this system. These data suggest that shear stress selectively activates uptake of L-arginine in endothelial cells and that the uptake of L-arginine might be important for shear stress-mediated nitric oxide formation.     

Bioaugmentation as a tool for improving the start-up and stability of a pilot-scale partial nitrification biofilm airlift reactor

The effectiveness of bioaugmentation in the improvement of the start-up of a biofilm airlift reactor to perform partial nitrification was investigated. Two identical biofilm airlift reactors were inoculated. The non-bioaugmented reactor (NB-reactor) was inoculated with conventional activated sludge, whereas the bioaugmented reactor (B-reactor) was seeded with the same conventional activated sludge but bioaugmented with nitrifying activated sludge from a pilot plant performing full nitritation under stable conditions (100% oxidation of influent ammonium to nitrite). The fraction of specialized nitrifying activated sludge in the inoculum of the B-reactor was only 6% (measured as dry matter). To simplify comparison of the results, operational parameters were equivalent for both reactors. Partial nitrification was achieved significantly faster in the B-reactor, showing a very stable operation. The results obtained by fluorescence in situ hybridization assays showed that the specialized nitrifying biomass added to the B-reactor remained in the biofilm throughout the start-up period.     

Disulfide bond formation contributes to herpes simplex virus capsid stability and retention of pentons

Disulfide bonds reportedly stabilize the capsids of several viruses, including papillomavirus, polyomavirus, and simian virus 40, and have been detected in herpes simplex virus (HSV) capsids. In this study, we show that in mature HSV-1 virions, capsid proteins VP5, VP23, VP19C, UL17, and UL25 participate in covalent cross-links, and that these are susceptible to dithiothreitol (DTT). In addition, several tegument proteins were found in high-molecular-weight complexes, including VP22, UL36, and UL37. Cross-linked capsid complexes can be detected in virions isolated in the presence and absence of N-ethylmaleimide (NEM), a chemical that reacts irreversibly with free cysteines to block disulfide formation. Intracellular capsids isolated in the absence of NEM contain disulfide cross-linked species; however, intracellular capsids isolated from cells pretreated with NEM did not. Thus, the free cysteines in intracellular capsids appear to be positioned such that disulfide bond formation can occur readily if they are exposed to an oxidizing environment. These results indicate that disulfide cross-links are normally present in extracellular virions but not in intracellular capsids. Interestingly, intracellular capsids isolated in the presence of NEM are unstable; B and C capsids are converted to a novel form that resembles A capsids, indicating that scaffold and DNA are lost. Furthermore, these capsids also have lost pentons and peripentonal triplexes as visualized by cryoelectron microscopy. These data indicate that capsid stability, and especially the retention of pentons, is regulated by the formation of disulfide bonds in the capsid.     

Microbiology and performance of a methanogenic biofilm reactor during the start-up period

Aims:  To understand the interactions between anaerobic biofilm development and process performances during the start-up period of methanogenic biofilm reactor.
Methods and Results:  Two methanogenic inverse turbulent bed reactors have been started and monitored for 81 days. Biofilm development (adhesion, growth, population dynamic) and characteristics (biodiversity, structure) were investigated using molecular tools (PCR–SSCP, FISH-CSLM). Identification of the dominant populations, in relation to process performances and to the present knowledge of their metabolic activities, was used to propose a global scheme of the degradation routes involved. The inoculum, which determines the microbial species present in the biofilm influences bioreactor performances during the start-up period. FISH observations revealed a homogeneous distribution of the Archaea and bacterial populations inside the biofilm.
Conclusion:  This study points out the link between biodiversity, functional stability and methanogenic process performances during start-up of anaerobic biofilm reactor. It shows that inoculum and substrate composition greatly influence biodiversity, physiology and structure of the biofilm.
Significance and Impact of the Study:  The combination of molecular techniques associated to a biochemical engineering approach is useful to get relevant information on the microbiology of a methanogenic growing biofilm, in relation with the start-up of the process.     

Biomass retention in a glucose acidifying anaerobic gas-lift reactor: isolation of the organism responsible for granule formation

Summary A study was undertaken of the microbial composition of aggregates from an acidifying anaerobic gas-lift reactor. For this purpose a simple 100 ml anaerobic gas lift reactor was developed. It was found that the predominant organism in the aggregates was Selenomonas ruminantium. Both, microscopical observations and a newly developed enumeration technique led to the conclusion that the mixed granules consisted mainly of this organism. Grown in pure culture, S. ruminantium was capable of forming aggregates. These aggregates resembled the mixed aggregates both macro- as well as microscopically. Furthermore the fermentation pattern of this pure aggregated culture was similar to that of a mixed aggregated culture.     

Diversity of ammonium-oxidizing bacteria in a granular sludge anaerobic ammonium-oxidizing (anammox) reactor

The ammonium-oxidizing microbial community was investigated in a granular sludge anaerobic ammonium-oxidizing (anammox) reactor that was operated for about 1 year with high anaerobic ammonium oxidation activity (up to 0.8 kg NH(4)(+)-N m(-3) day(-1)). A Planctomycetales-specific 16S rRNA gene library was constructed to analyse the diversity of the anaerobic ammonium-oxidizing bacteria (AnAOB). Most of the specifically amplified sequences (15/16) were similar to each other (> 99%) but were distantly related to all of the previously recognized sequences (< 94%), with the exception of an unclassified anammox-related clone, KSU-1 (98%). An ammonia monooxygenase (amoA) gene library was also analysed to investigate the diversity of 'aerobic' ammonium-oxidizing bacteria (AAOB) from the beta-Proteobacteria. Most of the amoA gene fragments (53/55) clustered in the Nitrosomonas europaea-Nitrosococcus mobilis group which has been reported to prevail under oxygen-limiting conditions. The quantitative results from real-time polymerase chain reaction (PCR) amplification showed that the dominant AnAOB comprised approximately 50% of the total bacterial 16S rRNA genes in the reactor, whereas the AAOB of beta-Proteobacteria represented only about 3%. A large fragment (4008 bp) of the rRNA gene cluster of the dominant AnAOB (AS-1) in this reactor sludge was sequenced and compared with sequences of other Planctomycetales including four anammox-related candidate genera. The partial sequence of hydrazine-oxidizing enzyme (hzo) of dominant AnAOB was also identified using new designed primers. Based on this analysis, we propose to tentatively name this new AnAOB Candidatus'Jettenia asiatica'.     

Fine-scale environmental variation contributes to introgression in a three-species spruce hybrid complex

Hybridization is common for many forest trees, where weak barriers to reproduction obscure species boundaries. We characterized the genomic structure of Picea populations comprising three species spanning two well-known contact zones, the Picea sitchensis?×?Picea glauca and the P. engelmannii?×?P. glauca hybrid zones, using a set of 71 candidate-gene single nucleotide polymorphisms. The genetic structure of populations suggests a complex genomic architecture shaped by interspecific gene flow and strong environmental selection, with increased genetic diversity in hybrids. The presence of admixture among all three species suggests that three-way hybrids with mixed ancestry occur where species ranges overlap in transitional environments. Significant clinal variation and associations with climatic variables (including continentality, temperature, and precipitation) differ between hybrid zones, indicating that individual species and their hybrids are adapted to distinct environmental niches. Allele–environmental association analysis revealed that most of the candidate genes with evidence of selection were unique to either the Sitka?×?white or the Engelmann?×?white hybrid zones, with few shared between these zones. Management of these widespread and diverse gene pools will be best served through development of climate-based seed transfer, with recommended seed sources informed by a combination of genetic and climatic information for future climates.     

Regime shift and microbial dynamics in a sequencing batch reactor for nitrification and anammox treatment of urine

The microbial population and physicochemical process parameters of a sequencing batch reactor for nitrogen removal from urine were monitored over a 1.5-year period. Microbial community fingerprinting (automated ribosomal intergenic spacer analysis), 16S rRNA gene sequencing, and quantitative PCR on nitrogen cycle functional groups were used to characterize the microbial population. The reactor combined nitrification (ammonium oxidation)/anammox with organoheterotrophic denitrification. The nitrogen elimination rate initially increased by 400%, followed by an extended period of performance degradation. This phase was characterized by accumulation of nitrite and nitrous oxide, reduced anammox activity, and a different but stable microbial community. Outwashing of anammox bacteria or their inhibition by oxygen or nitrite was insufficient to explain reactor behavior. Multiple lines of evidence, e.g., regime-shift analysis of chemical and physical parameters and cluster and ordination analysis of the microbial community, indicated that the system had experienced a rapid transition to a new stable state that led to the observed inferior process rates. The events in the reactor can thus be interpreted to be an ecological regime shift. Constrained ordination indicated that the pH set point controlling cycle duration, temperature, airflow rate, and the release of nitric and nitrous oxides controlled the primarily heterotrophic microbial community. We show that by combining chemical and physical measurements, microbial community analysis and ecological theory allowed extraction of useful information about the causes and dynamics of the observed process instability.     

Chitin contributes to the formation of a feeding structure in a predatory nematode

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Aerobic granulation and nitrogen removal with the effluent of internal circulation reactor in start-up of a pilot-scale sequencing batch reactor

Aerobic granular sludge was successfully cultivated with the effluent of internal circulation (IC) reactor in a pilot-scale sequencing batch reactor (SBR) using activated sludge as seeding sludge. N removal was investigated in the start-up of aerobic granulation process. Initially, the phenomenon of partial nitrification was observed and nitrite accumulation rates (NO₂ ^?-N/NO _x ^? -N) were between 84.6 and 99.1?%. It was potentially caused by ammonium oxidizing bacteria (AOB) in the seeding activated sludge, high external environmental temperature (~32?°C) and free ammonia (FA) concentration. After 50?days’ running, the aerobic granules-based bioreactor demonstrated perfect performance in simultaneous removal of organic matter and ammonia nitrogen, and average removal efficiencies were maintained above 93 and 96?%, respectively. The maximum nitrogen removal efficiency of 83.1?% was achieved after the formation of aerobic granules. The average diameter of mature aerobic granular sludge mostly ranged from 0.5 to 1.0?mm. Furthermore, one typical cyclic test indicated that pH and DO profiles could be used as effective parameters for biological reactions occurring in the aerobic/anoxic process. The obtained results could provide further information on the cultivation of aerobic granular sludge with practical wastewater, especially with regard to nitrogen-rich industrial wastewater.     

Asynchronous local dynamics contributes to stability of a seagrass bed in Tokyo Bay

It is known that asynchronous temporal variations in local populations can contribute to the stability of metapopulations. However, studies evaluating the hierarchical organization of multiple spatial scales are rare for continuous marine landscapes, especially for marine vegetation such as seagrass beds. In this study, long‐term observation (26 yr) of temporal changes and nested spatial analyses were combined for an extensive seagrass meadow in Tokyo Bay, Japan, using remote sensing and geographic information system technologies. We examined how the dynamics at the whole‐bed scale (~1 km²) are related to those at a local scale (0.04 km²), and investigated the relationship between the seagrass dynamics and long‐term changes in environmental conditions using data on oceanography, water quality, and sediment dynamics. The seagrass bed size fluctuated between a maximum of 1.28 km² (in 1987) and a minimum of 0.39 km² (in 2001), with an average of 0.90 km². The temporal variation in seagrass bed size at the whole‐bed scale correlated with sand movement within the seagrass bed related to changes in the position of a sandbar. Seagrass bed size fluctuated asynchronously at a local scale. Multivariate analyses recognized clusters of local areas showing similar patterns of fluctuation. Temporal patterns in the various clusters responded differently to changes in environmental factors, e.g. the position of the sandbar was highly correlated with seagrass bed size in shallow habitats but not in deeper areas. The magnitudes of the temporal variations for the local clusters were greater than that of the entire bed, suggesting that asynchronous fluctuation in different areas of the bed plays an important role in the overall stability of the seagrass bed. The results of the present study also highlight the importance of physical processes in regulating the temporal dynamics of seagrass beds in shallow sedimentary landscapes.     

Palmitoylation of LAT contributes to its subcellular localization and stability

Palmitoylation is a protein modification for trafficking to lipid raft. Without palmitoylation, linker for activation of T cells (LAT), an adaptor molecule mediating T cell receptor signaling, is unable to localize in lipid rafts and to mediate T cell activation. We here show a novel role for palmitoylation in LAT trafficking to the plasma membrane and in the stability of the LAT protein. The human LAT mutant lacking palmitoylation was unable to traffic to the plasma membrane despite the presence of transmembrane portion. The mouse LAT mutant lacking palmitoylation was unstable and susceptible to degradation via the proteasome pathway. The human LAT mutant became unstable when the extracellular portion was swapped for that from mouse, indicating that both palmitoylation and the extracellular portion regulate the stability of LAT. These results suggest that palmitoylation has an important role in trafficking to the plasma membrane and the stability of LAT.     

Granulation during the start-up of a UASB reactor used in the treatment of low strength wastewaters

A glucose-based wastewater was efficiently degradaded by acidogenic bacteria, with a glucose removal efficiency close to 90%, and although a distinctive granular structure could not be observed, fluffy conglomerates developed in an Upflow Anaerobic Sludge Blanket (UASB) reactor. Subsequently, the pre-acidification of the wastewater promoted the granulation process. An enrichment in methanogenic bacteria was observed on the microscope and was confirmed by an increase in the specific methanogenic activity from 0.1 up to 0.5kgCOD/kgVSS.day. Such dynamics of microbial communities was also verified through changes in the polysaccharide and protein content, as well as in the electrophoretic mobility of the biomass.