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PubMed2003

Diversity of nitrifying bacteria in full-scale chloraminated distribution systems.

Chloramination for secondary disinfection of drinking water often promotes the growth of nitrifying bacteria in the distribution system due to the ammonia introduced by chloramine formation and decay. This study involved the application of molecular biology techniques to explore the types of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) present in several full-scale chloraminated systems. The results of AOB community characterization indicated the ubiquitous detection of representatives from the Nitrosomonas genus, with Nitrosospira constituting a negligible or small fraction of the AOB community in all but one sample. Cloning and sequencing demonstrated the presence of AOB representatives within the Nitrosomonas oligotropha cluster, a phylogenetic subgroup of AOB from which isolates demonstrate a high affinity for ammonia. For the NOB communities, Nitrospira were detected in most of the samples, while Nitrobacter were only detected in a few samples. These results provide insight into the types of AOB responsible for nitrification episodes in full-scale chloraminated systems, which should help direct future studies aimed at characterizing relevant AOB growth and inactivation properties. Furthermore, the detection of NOB in most of the samples suggests a need to evaluate the contribution of biological nitrite oxidation relative to chemical oxidation in these systems.

Ammonia↗
PubMed2003

Dynamic response of nitrifying activated sludge batch culture to increased chloride concentration.

Dynamic response of nitrifying activated sludge batch cultures to increased chloride concentration was studied in this paper, which focused upon the changes in the specific nitrification rate (SNR) and nitrifier population when the chloride level was gradually or stepwise increased to 30,000 mg Cl L-1. The dominant species of ammonia-oxidizers and nitriteoxidizers in the population were examined by Fluorescent in situ hybridization technique with 16S rRNA-targeted oligonucleotide probes. It was found that neither chloride increasing approaches affected the SNR of the batch cultures before the chloride concentration exceeded 10,000 mg Cl L-1, after which the stepwise increase approach reduced the SNR more significantly than the gradual increase approach. From 10,000 to 18,000 mg Cl L-1 a down-and-up pattern of the SNR variation appeared in both approaches, which was associated with the change in the dominant species of ammonia-oxidizers from non-saline-resistant species such as Nitrosomonas europaea-lineage and Nitrosomonas eutropha to saline-resistant species, such as the Nitrosococcus mobilis-lineage. Nitrobacter was the only dominant species when the chloride concentration was below 10,000 mg Cl L-1, where no nitrite-oxidizers survived. Therefore, the 10,000 mg Cl L-1 chloride level is a critical level for the shift of the nitrifier population in the nitrifying activated sludge batch cultures.

Bioreactors↗
PubMed2001

Activity monitoring for nitrifying bacteria by fluorescence in situ hybridization and respirometry.

The activities of two families of nitrifying bacteria were measured by applied respirometer and fluorescence in situ hybridization (FISH) for the purpose of monitoring nitrogen removal. Oxygen uptake rates (OUR) for Nitrosomonas and Nitrobacter were separately measured with the dosing of selective inhibitors during respirometric monitoring. The FISH method was applied to analyze microorganism communities. In 5 mg NH4+-N/L solution, the total nitrogen oxygen demand (NOD) was found to be 6,672 mg DO/L-day with ammonia and nitrite oxidation rate of 4,512 and 2,160 mg DO/L-day, respectively. The oxygen consumption by endogenous oxidation was 288 mg DO/L-day and the biological oxygen demand 936 mg DO/L-day. Respirometric measurements were well matched by the FISH result, which described the presence and quantity of each nitrifying bacteria as nitrification proceeded. It was found that when coupled with OUR data, FISH could provide valuable information about the nitrogen removal process in many practical wastewater treatment systems.

Ammonia↗
PubMed2003

Structure and activity of multiple nitrifying bacterial populations co-existing in a biofilm.

A biofilm from a nitrifying pilot-scale sequencing batch reactor was investigated for effects of varying process conditions on its microscale activity and structure. Microsensor measurements of oxygen, substrates and products of nitrification were applied under incubation at different ammonium and oxygen concentrations which reflected various situations during a treatment cycle. A high net N loss was observed under high ammonium (HA) concentrations in contrast to low ones. Additionally, results indicated inhibition of nitrite-oxidizing bacteria (NOB), but not of ammonia-oxidizing bacteria (AOB) by free ammonia under HA conditions. Diversity, spatial distribution, and abundance of nitrifying bacteria as analysed by fluorescence in situ hybridization (FISH) revealed six different nitrifying populations with heterogeneous distributions. Nitrosococcus mobilis formed conspicuous microcolonies locally surrounded by cells of the dominating N. europaea/eutropha-related AOB population. A third less abundant population was affiliated to N. oligotropha. Nitrite-oxidizing bacteria of the genera Nitrobacter and Nitrospira (with at least two distinct populations) showed a large scale heterogeneity in their distribution. Nitrospira spp. were also found in deeper inactive layers where they might persist rather than thrive, and act as seed population when detached. Results of functional and structural analyses are discussed with respect to specific niches of individual populations in this system.

Ammonia↗
PubMed2001

The CANON system (Completely Autotrophic Nitrogen-removal Over Nitrite) under ammonium limitation: interaction and competition between three groups of bacteria.

The CANON system (Completely Autotrophic Nitrogen Removal Over Nitrite) can potentially remove ammonium from wastewater in a single, oxygen-limited treatment step. The usefulness of CANON as an industrial process will be determined by the ability of the system to recover from major disturbances in feed composition. The CANON process relies on the stable interaction between only two bacterial populations: Nitrosomonas-like aerobic and Planctomycete-like anaerobic ammonium oxidising bacteria. The effect of extended periods of ammonium limitation was investigated at the laboratory scale in two different reactor types (sequencing batch reactor and chemostat). The lower limit of effective and stable nitrogen removal to dinitrogen gas in the CANON system was 0.1 kg N m(-3) day(-1). At this loading rate, 92% of the total nitrogen was removed. After prolonged exposure (> 1 month) to influxes lower than this critical NH4+-influx, a third population of bacteria developed in the system and affected the CANON reaction stoichiometry, resulting in a temporary decrease in nitrogen removal from 92% to 57%. The third group of bacteria were identified by activity tests and qualititative FISH (Fluorescence In Situ Hybridisation) analysis to be nitrite-oxidising Nitrobacter and Nitrospira species. The changes caused by the NH4+-limitation were completely reversible, and the system re-established itself as soon as the ammonium limitation was removed. This study showed that CANON is a robust system for ammonium removal, enduring periods of up to one month of ammonium limitation without irreversible damage.

Bacteria, Anaerobic↗
PubMed2006

Selective enrichment and molecular characterization of a previously uncultured Nitrospira-like bacterium from activated sludge.

Previously uncultured nitrite-oxidizing bacteria affiliated to the genus Nitrospira have for the first time been successfully enriched from activated sludge from a municipal wastewater treatment plant. During the enrichment procedure, the abundance of the Nitrospira-like bacteria increased to approximately 86% of the total bacterial population. This high degree of purification was achieved by a novel enrichment protocol, which exploits physiological features of Nitrospira-like bacteria and includes the selective repression of coexisting Nitrobacter cells and heterotrophic contaminants by application of ampicillin in a final concentration of 50 microg ml(-1). The enrichment process was monitored by electron microscopy, fluorescence in situ hybridization (FISH) with rRNA-targeted probes and fatty acid profiling. Phylogenetic analysis of 16S rRNA gene sequences revealed that the enriched bacteria represent a novel Nitrospira species closely related to uncultured Nitrospira-like bacteria previously found in wastewater treatment plants and nitrifying bioreactors. The enriched strain is provisionally classified as 'Candidatus Nitrospira defluvii'.

Ampicillin↗
PubMed1977

Nitrification in histosols: a potential role for the heterotrophic nitrifier.

Insufficient populations of Nitrosomonas and Nitrobacter were found in a Pahokee muck soil (Lithic medidaprit) to account for the nitrate concentration observed. To determine if heterotrophic nitrifiers could account for some of this discrepancy, a method was developed to measure the levels of heterotrophic nitrifiers in soil. A population of 4.1 X 10(5) Arthrobacter per g of dry fallow soil, capable of producing nitrite and/or nitrate from reduced nitrogenous compounds, was observed. Amendment of the much with 0.5% (wt/wt) sodium acetate and 0.1% (wt/wt) ammonium-nitrogen as ammonium sulfate (final concentrations) not only resulted in the usual increase in autotrophic nitrifiers, but also in a fourfold increase in the heterotrophic nitrifying Arrthrobacter. Amendment of like samples with N-Serve [2-chloro-6(trichloromethyl) pyridinel] prevented the increase in Nitrosomonas, but not that in the heterotrophic nitrifiers. Nitrate production in the presence of the inhibitor was diminished but not prevented. An Arthrobacter sp., isolated from the muck, produced nitrite when inoculated at high densities into sterile soil, unamended or amended with sodium acetate and/or ammomium sulfate. These data suggest that the heterotrophic population may be responsible for some of the nitrate produced in these Histosols.

Acetates↗
PubMed1977

Nitrification in the intertidal zone: influence of effluent type and effect of tannin on nitrifiers.

Nitrification by intertidal sediments was measured by using a tide simulator that approximated the cycle of seawater on tidal flats. Sediments were chosen from sites affected by industrial and municipal effluents and pastoral seepage and runoff. The ability of sediments from different sites to nitrify endogenous nitrogen varied markedly. All sites exhibited an initial lag before activity commenced. The duration of this lag and the rate of nitrate production were different at each site. The sediments were also capable of oxidizing NH3-N supplied to them in seawater. This "nitrification potential" was highest at sites receiving nitrogenous effluents (slaughterhouse and sewage), but was also substantial in sediments affected by bark extract effluent and pasture runoff. The lowest potential and the longest lag were exhibited by sediments in an apple cannery effluent area. Enrichment cultures of nitrifying microorganisms were obtained from all sites using NH4+ as a source of energy, but enrichments for nitrite oxidizers were unsuccessful. Concentrated pine bark tannins, similar in origin to those in effluents at the well-nitrifying chipmill site, were tested for toxicity to pure cultures of nitrifying bacteria. Two Nitrobacter strains and one Nitrosomonas strain were unaffected by tannins even at 5 mg/ml. A Nitrosolobus and a Nitrosospira strain were inhibited partially at 5 mg/ml and only slightly or not at all at 1 mg/ml.

Ammonia↗
PubMed1996

Structure and function of a nitrifying biofilm as determined by in situ hybridization and the use of microelectrodes.

Microprofiles of O2 and NO3- were measured in nitrifying biofilms from the trickling filter of an aquaculture water recirculation system. By use of a newly developed biosensor for NO3-, it was possible to avoid conventional interference from other ions. Nitrification was restricted to a narrow zone of 50 microns on the very top of the film. In the same biofilms, the vertical distributions of members of the lithoautotrophic ammonia-oxidizing genus Nitrosomonas and of the nitrite-oxidizing genus Nitrobacter were investigated by applying fluorescence in situ hybridization of whole fixed cells with 16S rRNA-targeted oligonucleotide probes in combination with confocal laser-scanning microscopy. Ammonia oxidizers formed a dense layer of cell clusters in the upper part of the biofilm, whereas the nitrite oxidizers showed less-dense aggregates in close vicinity to the Nitrosomonas clusters. Both species were not restricted to the oxic zone of the biofilm but were also detected in substantially lower numbers in the anoxic layers and even occasionally at the bottom of the biofilm.

Biofilms↗
PubMed2003

Nitrification and nitrifying bacteria in the lower Seine River and estuary (France).

The Achères wastewater treatment plant, located just downstream of Paris, discharges its effluents into the lower Seine River. The effluents contain large numbers of heterotrophic bacteria, organic matter, and ammonium and are a source of nitrifying bacteria. As a result, degradation of organic matter by heterotrophic bacteria and subsequent oxygen depletion occur immediately downstream of the effluent outlet, whereas nitrifying bacteria apparently need to build up a significant biomass before ammonium oxidation significantly depletes the oxygen. We quantified the potential total nitrifying activity and the potential activities of the ammonia- and nitrite-oxidizing communities along the Seine River. In the summer, the maximum nitrifying activity occurs in the upper freshwater estuary, approximately 200 km downstream of Achères. The quantities of nitrifying bacteria, based on amoA gene copy numbers, and of Nitrobacter organisms, based on 16S rRNA gene copy numbers, were correlated with the potential nitrifying activities. The species composition of ammonia-oxidizing bacteria was investigated at two sites: the Triel station just downstream from Achères (km 84) and the Seine freshwater estuary at the Duclair station (km 278). By means of PCR primers targeting the amoA gene, a gene library was created. Phylogenetic analysis revealed that the majority of the analyzed clones at both sites were affiliated with the genus NITROSOMONAS: The Nitrosomonas oligotropha- and Nitrosomonas urea-related clones represented nearly 81% of the community of ammonia-oxidizing bacteria at Triel and 60% at Duclair. Two other ammonia-oxidizing clusters of the beta subclass of the Proteobacteria, i.e., Nitrosomonas europaea- and Nitrosospira-like bacteria, were found in smaller numbers. The major change in the ammonia-oxidizing community between the two stations along the Seine River-upper estuary continuum was the replacement of the N. oligotropha- and N. urea-related bacteria by the Nitrosospira-affiliated bacteria. Although the diversities of the ammonia oxidizers appear to be similar for the two sites, only half of the restriction patterns are common to both sites, which could be explained by the differences in ammonium concentrations, which are much lower in the upper estuary than in the river at the effluent outlet. These results imply a significant immigration and/or selection of the ammonia-oxidizing bacterial population along the continuum of the Seine River from Paris to the estuary.

Ammonia↗
PubMed2004

Dynamics of a pasture soil microbial community after deposition of cattle urine amended with [13C]urea.

Within grazed pastures, urine patches are hot spots of nitrogen turnover, since dietary N surpluses are excreted mainly as urea in the urine. This short-term experiment investigated 13C uptake in microbial lipids after simulated deposition of cattle urine at 10.0 and 17.1 g of urea C m(-2). Confined field plots without or with cattle urine amendment were sampled after 4 and 14 days, and soil from 0- to 5-cm and 10- to 20-cm depths was analyzed for content and composition of phospholipid fatty acids (PLFAs) and for the distribution of urea-derived 13C among individual PLFAs. Carbon dioxide emissions were quantified, and the contributions derived from urea were assessed. Initial changes in PLFA composition were greater at the lower level of urea, as revealed by a principal-component analysis. At the higher urea level, osmotic stress was indicated by the dynamics of cyclopropane fatty acids and branched-chain fatty acids. Incorporation of 13C from [13C]urea was low but significant, and the largest amounts of urea-derived C were found in common fatty acids (i.e., 16:0, 16:1omega7c, and 18:1omega7) that would be consistent with growth of typical NH4(+)-oxidizing (Nitrosomonas) and NO2(-)-oxidizing (Nitrobacter) bacteria. Surprisingly, a 20 per thousand depletion of 13C in the cyclopropane fatty acid cy17:0 was observed after 4 days, which was replaced by a 10 to 20 per thousand depletion of that in cy19:0 after 14 days. Possible reasons for this pattern are discussed. Autotrophic nitrifiers could not be implicated in urea hydrolysis to any large extent, but PLFA dynamics and the incorporation of urea-derived 13C in PLFAs indicated a response of nitrifiers which differed between the two urea concentrations.

Animals↗
PubMed1962

The effect of 2,3,6-trichlorophenylacetic and 2,2-dichloropropionic acids on nitrite oxidation.

The effects of two herbicides, 2,3,6-trichlorophenylacetic acid, sodium salt, and 2,2-dichloropropionic acid, sodium salt, on nitrite-oxidizing bacteria were studied by the soil perfusion technique. The time of application of 2,3,6-trichlorophenylacetic acid affected its toxicity to the nitrifier. When it was present in the environment as the nitrifier started growth, it was more toxic than if the organisms were allowed to nitrify actively before they were subjected to the herbicide. The herbicide 2,2-dichloropropionic acid at rates up to 700 ppm had little effect on nitrite oxidation. The toxicity of 2,3,6-trichlorophenylacetic acid for Nitrobacter was reduced by 2,2-dichloropropionic acid irrespective of whether the cells came into contact with the agents before or during active oxidation. The mode of action for this phenomenon has not been determined.

Bacteria↗
PubMed1967

Oxygen-nitrogen relationships in autotrophic nitrification.

Oxygen utilization by the autotrophic nitrifiers Nitrosomonas and Nitrobacter was studied. Experimental evidence is presented which reflects the effect of carbon dioxide fixation on overall oxygen utilization in autotrophic nitrification. Measurement of dissolved oxygen and inorganic nitrogen changes indicates that oxygen-nitrogen ratios in inorganic nitrogen oxidation are equal to 3.22 parts (expressed in milligrams per liter) of oxygen per part of ammonia nitrogen oxidized to nitrite nitrogen and 1.11 parts of oxygen per part of nitrite nitrogen oxidized to nitrate nitrogen. These values rather than the stoichiometric ratios should be used in nitrogenous oxygen demand calculations.

Ammonia↗
PubMed1971

Comparison of the morphology and deoxyribonucleic acid composition of 27 strains of nitrifying bacteria.

The gross morphology, fine structure, and per cent guanine plus cytosine (GC) composition of deoxyribonucleic acid of 27 strains of nitrifying bacteria were compared. Based on morphological differences, the ammonia-oxidizing bacteria were separated into four genera. Nitrosomonas species and Nitrosocystis species formed one homogenous group, and Nitrosolobus species and Nitrosospira species formed a second homogenous group in respect to their deoxyribonucleic acid GC compositions. Similarly, the nitrite-oxidizing bacteria were separated into three genera based on their morphology. The members of two of these nitrite-oxidizing genera, Nitrobacter and Nitrococcus, had similar GC compositions, but Nitrospina gracilis had a significantly lower GC composition than the members of the other two genera.

Ammonia↗
PubMed1972

Growth of obligate autotrophic bacteria on glucose in a continuous flow-through apparatus.

Nitrosomonas europaea, Nitrobacter agilis, Thiobacillus denitrificans, T. neapolitanus, and T. thioparus (all obligate autotrophic bacteria) have been grown in dialysis culture, on glucose salts media, in the absence of their specific inorganic energy source. Metabolic products for N. agilis grown on nitrite salts medium were identified as keto acids. Pyruvic acid inhibited this organism at 5 x 10(-5)m. Keto acids were not inhibitory for the thiobacilli grown on thiosulfate medium. However, when T. denitrificans was grown on glucose with dialysis, addition of 5 x 10(-4)m pyruvate inhibited growth. Thus, it appears pyruvate may be inhibitory for other autotrophs, as has been reported for T. thiooxidans.

Ammonia↗
PubMed1975

Nitrification on a coral reef.

We report that the algal pavement just behind the reef crest at Enewetak Atoll produces nitrate at measurable rates. In situ and in vitro incubations with N-Serve indicate that the autotrophic pathway involving two separate organisms is effective in this oxidation of ammonia to nitrate. Significant nitrification is indicated throughout the reef environment; Nitrobacter agilis has specifically been identified as at least one of the organisms responsible for the terminal oxidation of nitrite to nitrate.

Ammonia↗
PubMedDate not supplied

Influence of chemical oxygen demand/total Kjeldahl nitrogen ratio and sludge age on nitrification of nitrogenous wastewater.

Four laboratory-scale biological nitrification units (influent total Kjeldahl nitrogen [TKN] = 1002 to 1062 mg/L) were operated at chemical oxygen demand (COD)/TKN ratios of approximately 0.5, 1.0,15, and 2.0 and at three different sludge ages of 30, 20, and 10 days to study the influence of COD/TKN, sludge age, COD loading, and TKN loading on nitrification and nitrifiers. Percent nitrification was found to increase with decreases in COD/TKN and increases in sludge age. The average nitrifier concentration increased from 460 mg/L at a COD/TKN of 2.22 and a sludge age of 10 days to 706 mg/L at a COD/TKN of 0.676 and a sludge age of 30 days. The nitrifier fraction was found to be higher at a lower COD/TKN and lower at a higher COD/TKN. The nitrifier fraction increased with the decrease in sludge age and COD loadings and the increase in TKN loadings. The effect of sludge age on the nitrifier fraction was amplified at a COD/ TKN of approximately 0.5 rather than at approximately 2.0. The nitrification rate (kilograms TKN oxidized per kilograms nitrifiers per day) was shown to be dependent on COD/TKN and sludge age. The activity performed by Nitrobacter was affected at all COD/TKN ratios studied as well as at a sludge age of 10 days. This was manifested by the accumulation of high levels of nitrite-nitrogen in the nitrified effluent. The presence of heterotrophs did not affect nitrification rates and the growth of nitrifiers, which were found to be beneficial. High sludge age and COD loadings resulted in a higher sludge volume index of more than 200 mL/g mixed liquor suspended solids. Microscopic examination showed filamentous structure of sludge under these conditions. It is concluded from the investigations that a sludge age of 30 days and a COD/TKN of approximately 1.0 are optimal to yield maximum nitrification and nitrifier growth rates for treating high-strength nitrogenous wastewater.

Bioreactors↗
PubMed1999

Water quality management for low temperature marine fishes in space.

Vestibular Function Experiment Unit (VFEU), one of the Spacelab facility flown in Neurolab mission (STS-90) in April, 1998, was to support neurophysiological research using a marine fish, Opsanus tau (oyster toadfish). The functions of the VFEU were primarily a quality management of environmental water during the mission at 14 degrees C and for acquiring physiological signals from implanted micro-electrodes in the otolith nerves as well as the spatial acceleration of the fish. A key element of the life support system was a balanced biological filter containing two types of nitrifying bacteria, Nitrosomonas for ammonia oxidization and Nitrobacter for nitrite oxidization. Although the 16 days mission was successful, two toadfishes died in late phase of the mission. Ammonium concentration in those two life support systems elevated to remarkably high level at the end of the mission whereas the other two indicated very low. This report summarizes the results of the water quality management of the VFEU during the Neurolab mission based on analysis of water samples taken during the flight and those taken prior and just after the flight.

Animals↗