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Implementation, study and calibration of a modified ASM2d for the simulation of SBR processes.

An enhanced process model for SBRs has been developed. Though the basic mechanism largely draws on the Activated Sludge Model n. 2d, its new features are the splitting of the nitrification stage in a two-step process, according to the well known Nitrosomonas-Nitrobacter oxidation sequence, and an improved XPAO dynamics, involved in the anaerobic/aerobic phosphorus removal process. The model was implemented through the DLL technique allowing complied C++ modules to be linked to an ordinary Simulink block diagram. The static sensitivity study revealed that if the parameter vector is partitioned into subsets of biologically related parameters and calibrated separately, the calibration procedure does not present particularly difficult aspects. Trajectory sensitivity showed also to which extent data collection could be optimised in order to improve calibration accuracy. The study of the shape of the error functional generated by parameters couples allows a much more effective calibration strategy.

Bioreactors↗

Go or no go for gel entrapped nitrifiers? A Belgian case study.

In the coming years, as stricter environmental requirements are imposed, many European Union wastewater treatment plants (WWTP) need to be expanded and/or upgraded. This requires considerable investments. Optimising the renovation recourses can lead to significant savings. The use of entrapped nitrifying bacteria for upgrading of WWTP towards nutrient removal may be benificial. Long term pilot tests were performed to evaluate a so-called pellet reactor. Differences in performance and microbiological composition of classical activated sludge and the pellet reactor were investigated. FISH analyses showed (i) absence of Nitrobacter cells and (ii) high abundance of Nitrospira in the pilot reactors. Two Belgian WWTP make use of fine bubble aeration and could--theoretically--easily be renovated towards nitrogen removal using encapsulated nitrifiers. Financial aspects are commented on.

Belgium↗

Nitrite accumulation followed by denitrification using sequencing batch reactor.

Biological ammonia-nitrogen removal utilizes two distinct processes, nitrification and denitrification. In nitrification, ammonia oxidizes to nitrite then to nitrate. In this study, elimination of nitrite oxidation to nitrate step was attempted in order to directly remove nitrite to nitrogen gas by denitrification. For this study the supernatant from an anaerobic digester was used as an ammonia source and a sequencing batch reactor (SBR) was employed. Emphasis was given to the evaluation of the operational factors affecting nitrite accumulation and the elucidation of kinetics for biological nitrification and denitrification. Accumulation of nitrite in the nitrification process was achieved by suppressing the growth of Nitrobacter, a nitrite oxidizer, by loading high concentration ammonia supernatant immediately after all ammonia in the previous loading was oxidized to nitrite. Nitrite oxidation was taking place as the solid retention time (SRT) was increased from 2.5 days to 3.0 days in a continuously aerated SBR mode with daily feeding. However, nitrite accumulation was achieved even at longer SRT of 5 days when the aeration and non-aeration periods were appropriately combined and the non-aeration period can be used for denitrification of the accumulated nitrite with a carbon source supplied.

Ammonia↗

Using a compact combined constructed wetland system to treat agricultural wastewater with high nitrogen.

The objectives of this study were to find appropriate conditions for nitrogen removal by a compact combined constructed wetland system and to evaluate the removal rate constant in a tropical climate. This study will present suitable operating conditions for a combined system to treat pig farm wastewater containing high ammonia-nitrogen. Four laboratory-scale combined constructed wetland units (0.5 x 1.0 x 1.0 m3): vertical flow vegetated bed over horizontal flow sand bed, were operated under an average temperature of 24 degrees C. Pig farm wastewater with COD and NH4-N concentration of 1034 and 448 mg/L in average was fed to the system at different HLR from 2 to 8 cm/day. The performance of the system when operated with a vertical flow bed followed by a horizontal flow bed or vice versa did not show a significant difference but under high HLR, nitrogen removal efficiencies were clearly reduced. Nitrobacter and Nitrosomonas were found in a large number in vertical flow beds and the same for denitrifier bacteria in a horizontal flow beds. Removal rate constants for nitrification (kNH4+ -N) were 0.0413 m/d for H-Vmode and 0.0339 m/d for V-H mode. Removal rate constants for denitrification (kNOx-N) were 0.0979 m/d for H-Vmode and 0.0399 m/d for V-H mode, respectively.

Agriculture↗

N-terminal amino acid sequence of cytochrome c-552 from Nitrosomonas europaea.

Nitrosomonas europaea is an ammonia-oxidizing bacterium which contains multiple c-type cytochromes. Few of these components have been assigned physiological roles, but on the basis of molecular weight and redox potential cytochrome c-552 has been considered to be an analogue of the mitochondrial cytochrome-c family of proteins. We present the N-terminal amino acid sequence (47 residues) of cytochrome c-552 and show that this protein is most closely related to the group of small cytochrome-c components from pseudomonads (cytochromes c-551) and is probably evolutionarily distant from the analagous protein (cytochrome c-550) from the nitrite-oxidizing bacterium Nitrobacter agilis.

Amino Acid Sequence↗

[Participation of nitrifying bacteria in the disintegration of serpentinous ultrabasic rock].

Nitrifying bacteria were found to be widely distributed among the products of the weathering crust of ultrabasite rocks. Nitrosospira briensis and Nitrobacter winogradskyi involved in the first and second phases of nitrification, respectively, were detected and isolated as pure cultures. In experiments conducted with a pure culture of Nitrosospira briensis, a correlation was established between degradation of serpentinite by this culture and an increase in the content of nitrites in the growth medium. The presence of nitrogen compounds in the deposits, as well as wide distribution of nitrifying bacteria, suggests that this bacterial group along with other, in particular, heterotrophic microorganisms participates in weathering of serpentinized ultrabasite rocks, leaching of elements, and formation of the weathering crust.

Bradyrhizobiaceae↗

[The effect of pH and partial pressure of oxygen on nitrification (author's transl)].

A mathematical model which describes the effect of pH and dissolved oxygen on growth and activity of Nitrosomonas and Nitrobacter in mixed culture is presented. A good fit is obtained between experimental data and theoretical curves computed with parameters estimated by independent methods. Experimental data together with simulation studies show that inadequate aeration induces a temporal shift of ammonium and nitrite oxidations resulting in a transient nitrite accumulation similar to that caused by an increase of temperature.

Hydrogen-Ion Concentration↗

Effect of high concentrations of nitrogen on mixed populations of nitrifying bacteria isolated from industrial nitrogenous wastewaters.

The effect of high concentrations of different forms of nitrogen (NH4+, NO2-, NO3- and urea N) on nitrification by mixed populations of nitrosobacters isolated from nitrogen fertilizer plant wastewaters and nitrobacters isolated from effluents from a biological bed treating these wastewaters was determined. The inhibitory activity (within the concentration values for industrial wastewaters) of only the reaction products was observed, i.e. nitrites for nitrification phase I and nitrates for nitrification phase II. A mixed population of phase II nitrifying bacteria is highly resistant to high concentrations of ammonia nitrogen (50% inhibition at 3,000 mg N/l).

Ammonia↗

Nitrifying bacterial communities in an aquaculture wastewater treatment system using fluorescence in situ hybridization (FISH), 16S rRNA gene cloning, and phylogenetic analysis.

Aquaculture, especially shrimp farming, has played a major role in the growth of Thailand's economy in recent years, as well as in many South East Asian countries. However, the nutrient discharges from these activities have caused adverse impacts on the quality of the receiving waterways. In particular nitrogenous compounds, which may accumulate in aquaculture ponds, can be toxic to aquatic animals and cause environmental problems such as eutrophication. The mineralization process is well known, but certain aspects of the microbial ecology of nitrifiers, the microorganisms that convert ammonia to nitrate, are poorly understood. A previously reported enrichment of nitrifying bacteria (ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB)) from a shrimp farm inoculated in a sequencing batch reactor (SBR) was studied by molecular methods. The initial identification and partial quantification of the nitrifying bacteria (AOB and NOB) were carried out by fluorescence in situ hybridization (FISH) using previously published 16S rRNA-targeting oligonucleotide probes. The two dominant bacterial groups detected by FISH were from the Cytophaga-Flavobacterium-Bacteroides and Proteobacteria (beta subdivision) phyla. Published FISH probes for Nitrobacter and Nitrospira did not hybridize to any of the bacterial cells. Therefore it is likely that new communities of NOBs, differing from previously reported ones, exist in the enrichments. Molecular genetic techniques (cloning, sequencing, and phylogenetic analysis) targeting the 16S rRNA genes from the nitrifying enrichments were performed to identify putative AOBs and NOBs.

Animals↗

Taxon-specific content of oligonucleotide triplets in 16S rRNAs of anoxygenic phototrophic and nitrifying bacteria.

Theoretical evaluation of the content of oligonucleotide triplets AAA, CCC, and UAU in 16S rRNAs of anoxygenic phototrophic bacteria (genera Chlorobium; Chloroflexus; Chromatium: Rhodopseudomonas) and nitrifying bacteria (genera Nitrosococcus, Nitrosomonas, Nitrosolobus, Nitrosovibrio, Nitrospira, Nitrospina, Nitrobacter) showed that the number of the AAA, CCC or UAU triplets in 16S rRNAs specifically corresponds to the genus and species of bacteria. The ratio of AAA and CCC triplet numbers in the sequences of 16S rRNA (AAA/CCC) of anoxygenic phototrophic bacteria was within the range of 0.61 to 2.03, and the ratio of AAA and UAU (AAA/UAU) triplet numbers in the sequence of 16S rRNA was within the range of 2.88 to 12.00. The regions of any genus within the AAA/CCC and AAA/UAU axes did not overlap. The combination of the numbers of nucleotide triplets in 16S rRNA is genus-specific character. The similar data were obtained in the study of a physiological group of nitrifying bacteria. The range of AAA/UAU ratio was from 1.8 to 9.0, and range of AAA/CCC was from 0.9 to 2.6 for this taxon. The number of triplets in 16S rRNAs of the studied taxa was genus- and species-specific character. The biological significance of these data is the evidence that not only the sequence but the number of nucleotide triplets in 16S rRNAs reflects the phylogeny of corresponding taxa.

Bacteria↗

Biotreatment of ammonia- and butanal-containing waste gases.

The biological removal of ammonia and butanal in contaminated air was investigated by using, respectively, a laboratory-scale filter and a scrubber-filter combination. It was shown that ammonia can be removed with an elimination efficiency of 83% at a volumetric load of 100 m3.m-2.h-1 with 4-16 ppm of ammonia. During the experiment percolates were analysed for nitrate, nitrite, ammonium and pH. It was found that the nitrification in the biofilter could deteriorate due to an inhibition of Nitrobacter species, when the free ammonia concentration was rising in the percolate. It should be easy to control such inhibition through periodic analysis of the liquid phase by using a filter-scrubber combination. Such a combination was studied for butanal removal. Butanal was removed with an elimination efficiency of 80% by a scrubber-filter combination at a volumetric load of 100 m3.m-2.h-1 and a high butanal input concentration. Mixing the filter material with CaCO3 and pH control of the liquid in the scrubber resulted in an increase of the elimination efficiency. These results, combined with previous results on the biofiltration of butanal and butyric acid, allow us to discuss the influence of odour compounds on the removal efficiency of such systems and methods for control. The results were used to construct a full-size system, which is described.

Air Pollutants↗

A phylogenetic survey of budding, and/or prosthecate, non-phototrophic eubacteria: membership of Hyphomicrobium, Hyphomonas, Pedomicrobium, Filomicrobium, Caulobacter and "dichotomicrobium" to the alpha-subdivision of purple non-sulfur bacteria.

The phylogenetic position of various budding and/or or prosthecate Gram-negative eubacteria was determined by different methods. Members of the genera Hyphomicrobium, Filomicrobium, Pedomicrobium were investigated by 16S rRNA cataloguing, a 1373 nucleotide long portion of the 16S rRNA was sequenced from Hyphomicrobium vulgare and the 5S rRNAs were analyzed from two Hyphomicrobium strains, Hyphomonas polymorpha and Caulobacter crescentus. Comparison with published sequences indicated a membership of all of these organisms to the alpha subdivision of purple bacteria. While C. crescentus and Hyphomonas polymorpha constitute separate individual lines of descent, the position of a coherent cluster embracing Hyphomicrobium, Pedomicrobium and Filomicrobium is not yet settled. 16S rRNA cataloguing indicate the presence of a distinct line equivalent to other subgroups in its phylogenetic depth. 5S rRNA analysis, on the other hand, groups Hyphomicrobium vulgare and strain IFAM 1761 with members of subgroup alpha-2 (Rhodopseudomonas palustris, Nitrobacter winogradskyi and relatives). In contrast to the present classification, Pedomicrobium ferrugineum and Filimicrobium fusiforme are more closely related to certain Hyphomicrobium strains than these are related among each other. Budding mode of reproduction and prosthecate morphology are dominating morphological features of members of the alpha subdivision. These characteristics may gain diagnostic significance in a future formal description of this subdivision and its subgroups as a higher rank.

Base Sequence↗

A new obligately chemolithoautotrophic, nitrite-oxidizing bacterium, Nitrospira moscoviensis sp. nov. and its phylogenetic relationship.

A gram-negative, non-motile, non-marine, nitrite-oxidizing bacterium was isolated from an enrichment culture initiated with a sample from a partially corroded area of an iron pipe of a heating system in Moscow, Russia. The cells were 0.9-2.2 microns x 0.2-0.4 microns in size. They were helical- to vibroid-shaped and often formed spirals with up to three turns 0.8-1.0 micron in width. The organism possessed an enlarged periplasmic space and lacked intracytoplasmic membranes and carboxysomes. The cells tended to excrete extracellular polymers, forming aggregates. The bacterium grew optimally at 39 degrees C and pH 7.6-8.0 in a mineral medium with nitrite as sole energy source and carbon dioxide as sole carbon source. The optimal nitrite concentration was 0.35 mM. Nitrite was oxidized to nitrate stoichiometrically. The doubling time was 12 h in a mineral medium with 7.5 mM nitrite. The cell yield was low; only 0.9 mg protein/l was formed during oxidation of 7.5 mM nitrite. Under anoxic conditions, hydrogen was used as electron donor with nitrate as electron acceptor. Organic matter (yeast extract, meat extract, peptone) supported neither mixotrophic nor heterotrophic growth. At concentrations as low as 0.75 g organic matter/l or higher, growth of nitrite-oxidizing cells was inhibited. The cells contained cytochromes of the b- and c-type. The G+C content of DNA was 56.9 +/- 0.4 mol%. The chemolithoautotrophic nitrite-oxidizer differed from the terrestrial members of the genus Nitrobacter with regard to morphology and substrate range and equaled Nitrospira marina in both characteristics. The isolated bacterium is designated as a new species of the genus Nitrospira.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacterial Proteins↗

Organic halogen removal from chlorinated humic ground water and lake water by nitrifying fluidized-bed biomass characterised by electron microscopy and molecular methods.

The dechlorinating and genotoxicity-removing activities of nitrifying fluidized-bed reactor biomass towards chlorinated organic compounds in water were shown at level below 1 ppm. The removal rates of adsorbable organic halogens were 200 micrograms Cl (g VS day)-1 for chlorinated humic ground water and 50 micrograms Cl (g VS day)-1 for chlorinated lake water when studied in batch mode. In a sequenced batch mode the removal rates [micrograms Cl (g VS day)-1] were 2000 from chlorohumus, 1400-1800 from chlorophenols in chlorinated ground water, and 430-720 from chlorohumus in chlorinated lake water. Genotoxicity was removed to a large extent (60%-80%) from the chlorinated waters upon incubation with nitrifying reactor biomass. 2,6-Di-, 2,4,6-tri and 2,3,4,6-tetrachlorophenols competed with chlorinated water organohalogens for dechlorination. The dechlorination of chlorophenols and chlorohumus required no ammonia and was not prevented by inhibitors of ammonia oxidation, nitrapyrin, parathion, sodium diethyldithiocarbamate, or allylthiourea. Electron microscopical inspection of the biomass showed the dominance of clusters of bacteria resembling known nitrifying species, Nitrosomonas, Nitrobacter, and Nitrosospira. This was supported by polymerase chain reaction amplification of the biomass DNA with four different primers, revealing the presence of 16S rDNA sequences assignable to the same species. The most intensive band obtained with the Nitroso4E primer was shown to be closely related to Nitrosomonas europaea by restriction analysis.

Ammonia↗

An analysis of nitrification during the aerobic digestion of secondary sludges.

Investigations were undertaken to study the occurrence and progress of nitrification during aerobic digestion of activated sludge in a wide range of initial concentrations of total solids (1000 to 80 000 mg litre(-1), initial pH range of 4.5 to 10.4 and digestion temperature range of 5 degrees to 60 degrees C. Batch aerobic digestion studies on activated sludge grown on wastewater (enriched with organic solids from human excretal material) indicate that almost complete elimination of the 'biodegradable' matter of the activated sludge was one of the essential prerequisites to initiate nitrification. Favourable ranges of temperature and pH for nitrification were observed to be 25 degrees to 30 degrees C and 6.0 to 8.3, respectively. With all favourable conditions, a minimum period of about 2 days was necessary for population build-up of genera Nitrosomonas and Nitrobacter, and to initiate nitrification. Nitrate formation invariably lagged behind nitrite formation, but under certain conditions both phases of nitrification were observed to progress hand in hand.

Journal Article↗

Modeling microbial-mediated reduction in batch reactors.

The governing equations that depict microbially-mediated reduction of heavy metals in the subsurface include a system of coupled nonlinear partial differential equations (PDE's) that describe physical (transport), chemical (sorption), and microbial (reduction/oxidation) processes. The existence of nonlinear reaction terms makes numerical simulations more challenging; however, with the advent of time-splitting solution algorithms, nonlinear reaction terms can be isolated from the convective-dispersive components of the governing transport equations and then solved as a coupled system of nonlinear ordinary differential equations (ODE's). In this paper, four methods are evaluated for solving coupled systems of nonlinear ODE's that describe microbially-mediated reduction/oxidation processes. The evaluation involves a series of comparisons of transient simulations of electron donor oxidation, electron acceptor reduction, and microbial biomass accumulation. The methods evaluation is initiated with a comparison of simulation results obtained with the four methods to those generated with an analytical model. Next, laboratory observations, of nitrite consumption by Nitrobacter winogradski in batch reactors are used in a comparison of batch system simulations generated using each of the four methods and BIOKEMOD (biogeochemical kinetic/equilibrium reaction model). The evaluation finds one of the four methods, the quasi-steady-state approximation (QSSA), to be among the most accurate and easiest to implement. Final validation of the QSSA is performed simulating experimental results of microbially-mediated chromium reductions in batch cultures.

Bacteria↗

Chemolithotrophic haloalkaliphiles from soda lakes.

This paper summarizes recent data on the occurrence and properties of lithotrophic prokaryotes found in extremely alkaline, saline (soda) lakes. Among the chemolithotrophs found in these lakes the obligately autotrophic sulfur-oxidizing bacteria were the dominant, most diverse group, best adapted to haloalkaline conditions. The culturable forms are represented by three new genera, Thioalkalimicrobium, Thioalkalivibrio and Thioalkalispira in the Gammaproteobacteria. Among them, the genus Thioalkalivibrio was most metabolically diverse, including denitrifying, thiocyanate-oxidizing and facultatively alkaliphilic species. Culturable methane-oxidizing populations in the soda lakes belong to the type I methanotroph group in the Gammaproteobacteria, mostly in the genus Methylomicrobium. The nitrifying bacteria in hyposaline soda lakes were represented by a new species Nitrobacter alkalicus (Alphaproteobacteria), and by an alkaliphilic subspecies of Nitrosomonas halophila (Betaproteobacteria). Both belonged to the low salt-tolerant alkaliphiles. The facultatively autotrophic haloalkaliphilic isolates able to grow with hydrogen as electron donor were identified as representatives of the alpha-3 subclass of the Proteobacteria (aerobic) and of the Natronolimnicola - Alkalispirillum group in the gammaproteobacteria (nitrate-reducing). While all chemolithotrophic isolates from soda lakes belong to the alkaliphiles with a pH optimum for growth around 10, only the sulfur-oxidizing group included species able to grow under hypersaline conditions. This indicates that carbon and nitrogen cycles in the hypersaline alkaline lakes might not be closed.

Fresh Water↗

High levels of nitrifying bacteria in intermittently aerated reactors treating high ammonia wastewater.

Changes in the fractions of ammonia-oxidizing bacteria and nitrite-oxidizing bacteria in two laboratory-scale reactors were investigated using 16S rRNA probe hybridizations. The reactors were operated in intermittent aeration mode and different aeration cycles to treat anaerobically digested swine wastewater with ammonia concentrations up to 175 mg NH(3)-N/L. High ammonia removals (>98.8%) were achieved even with increased nitrogen loads and lower aeration: non-aeration time ratios of 1h:3h. Nitrosomonas/Nitrosococcus mobilis were the dominant ammonia-oxidizing bacteria in the reactors. Nitrospira-like organisms were the dominant nitrite-oxidizing bacteria during most of the investigation, but were occasionally outcompeted by Nitrobacter. High levels of nitrifiers were measured in the biomass of both reactors, and ammonia-oxidizing bacteria and nitrite-oxidizing bacterial levels adjusted to changing aeration: non-aeration time ratios. Theoretical ammonia-oxidizer fractions, determined by a mathematical model, were comparable to the measured values, although the measured biomass fractions were different at each stage while the theoretical values remained approximately constant. Stable ammonia removals and no nitrite accumulation were observed even when rRNA levels of ammonia oxidizers and nitrite-oxidizers reached a minimum of 7.2% and 8.6% of total rRNA, respectively. Stable nitrogen removal performance at an aeration: non-aeration ratio of 1h:3h suggests the possibility of significant savings in operational costs.

Ammonia↗