Changes in the bacterial community structure in two-stage constructed wetlands with different plants for industrial wastewater treatment

This study focused on the diversity of bacterial communities from two series of two-stage constructed wetlands (CWs) treating tannery wastewater, under different hydraulic conditions. Series were separately planted with Typha latifolia and Phragmites australis in expanded clay aggregates and operated for 31 months. The effect of plant species, hydraulic loading and unit stage on bacterial communities was addressed through bacterial enumeration and denaturating gradient gel electrophoresis (DGGE). Diverse and distinct bacterial communities were found in each system unit, which was related in part to the type of plant and stage position (first or second unit in the series). Numerical analysis of DGGE profiles showed high diversity in each unit with an even distribution of species. No clear relation was established between the sample collection time, hydraulic loading applied and the bacterial diversity.     

Study of microbial community structures in UASB sludge treating municipal wastewater by denaturing gradient gel electrophoresis of 16S rDNA

The structures of microbial communities in lab-scale upflow anaerobic sludge blanket (UASB) reactors for treating municipal wastewater with different ratios of COD_soluble/ COD_total were studied using denaturing gradient gel electrophoresis (DGGE) of 16S rRNA genes. The microbial structure of the inoculum sludge obtained from a full-scale UASB reactor of treating potato processing wastewater was compared with the structures of sludges collected from three lab-scale UASB reactors after eight months feeding with raw municipal wastewater, with CEPS (chemically enhanced primary sedimentation) pretreated municipal wastewater, and with a synthetic municipal sewage, respectively. Computer-aided numerical analysis of the DGGE fingerprints showed that the bacterial community underwent major changes. The sludges for treating raw and CEPS pretreated wastewater had very similar bacterial and archaeal communities (82% and 96% similarity) but were different from that for treating the synthetic sewage. Hence, despite similar % COD in the particulate form in the synthetic and the real wastewater, the two wastewaters were selected for different microbial communities. Prominent DGGE bands of Bacteria and Archaea were purified and sequenced. The 16S rRNA gene sequences of the dominant archaeal bands found in the inoculum, and UASB sludge fed with raw sewage, CEPS pretreated wastewater, and synthetic sewage were closely associated withMethanosaeta concilii. In the UASB sludge fed with synthetic sewage, another dominant band associated with an uncultured archaeon 39-2 was found together withM. concilii.     

Effect of monoculture soybean on soil microbial community in the Northeast China

Monoculture (MC) soybean, a common practice in the Northeast China, causes significant declines in soybean yield and quality. The objective of this study was to evaluate the responses of the soil microbial community and soybean yield to different soybean cropping systems. Three cropping systems were compared, (1) corn-soybean rotation (corn-corn-soybean, CS), (2) MC soybean for 3 years (S3), (3) MC soybean for 9 years (S9). Both bulk and rhizosphere soil samples were collected at three growth stages: two trifoliate (V2), full bloom (R2), and full seed (R6), respectively. Soil microbial DNA was analyzed using polymerase chain reaction (PCR)—denaturing gradient gel electrophoresis (DGGE) to assess changes in composition of bacterial and fungal communities. Prominent DGGE bands were excised and sequenced to gain insight into the identities of the predominant microbial populations. Some prominent differences were observed in bacterial DGGE patterns of amplified 16S rDNA (V3 region) among rhizosphere soils. These major differences included one DGGE band (showing 100% similarity to Arthrobacter sp.) that was enriched at R2 stages in CS and S9, and another band with 97% sequence similarity to an uncultured actinobacterium was detected at R6 stage in CS, and at R2 and R6 stages in S9. The bacterial community from bulk soil showed no significant band change in DGGE patterns among different cropping systems. In fungal DGGE patterns of the amplified 18S rDNA partial fragment, one specific band (showing 98% similarity to Trichoderma viride) occurred in rhizosphere soil of treatment CS at V2 and R6 stages and treatment S9 at R6 stage. None of the above bands were detected in treatment S3. The soybean yields and plant heights from CS and S9 were greater than those from S3. Moreover, catalase activities from CS and S9 at V2 and R2 stages were higher than those tested from S3 at the corresponding times in rhizosphere soil. The present results showed that DGGE patterns were not able to detect significant differences in diversity or evenness among microbial communities, but significant differences were found in the composition of bacterial and fungal community structures. Some distinguished bands from bacterial and fungal DGGE patterns were only enriched in CS and S9 soil, which could potentially play an important role in soybean growth development.     

Microbial ecology of autothermal thermophilic aerobic digester (ATAD) systems for treating waste activated sludge

Despite their widespread use, our understanding of the microbial ecology of the autothermal thermophilic aerobic digesters (ATAD) used to dispose of sludge from wastewater treatment plants is poor. Applying both culture-dependent and molecular methods to two ATAD systems in Victoria, Australia treating different wastewaters revealed that their communities were highly specialized. Denaturing gradient gel electrophoresis (DGGE) profiling suggested differences in their population compositions and both changed over time. However, both showed low level biodiversity, and contained several novel bacterial populations. 16S rRNA clone library data and FISH analyses showed that Thermus thermophilus dominated both communities and that of a third ATAD plant in NSW (more than 90% of the total bacterial biovolume in repeated samples taken from each of the three ATAD plants). Culture-dependent methods also showed Geobacillus spp. were present in both Victorian communities. Nevertheless, the ecophysiology of these populations and their putative roles in sludge digestion remain unclear. FISH/microautoradiographic studies did not provide conclusive data elucidating which substrate/s T. thermophilus might utilize in the ATAD reactors.     

Changes in Bacterial Communities of the Marine Sponge Mycale laxissima on Transfer into Aquaculture

The changes in bacterial communities associated with the marine sponge Mycale laxissima on transfer to aquaculture were studied using culture-based and molecular techniques. M. laxissima was maintained alive in flowthrough and closed recirculating aquaculture systems for 2 years and 1 year, respectively. The bacterial communities associated with wild and aquacultured sponges, as well as the surrounding water, were assessed using 16S rRNA gene clone library analysis and denaturing gradient gel electrophoresis (DGGE). Bacterial richness and diversity were measured using DOTUR computer software, and clone libraries were compared using S-LIBSHUFF. DGGE analysis revealed that the diversity of the bacterial community of M. laxissima increased when sponges were maintained in aquaculture and that bacterial communities associated with wild and aquacultured M. laxissima were markedly different than those of the corresponding surrounding water. Clone libraries of bacterial 16S rRNA from sponges confirmed that the bacterial communities changed during aquaculture. These communities were significantly different than those of seawater and aquarium water. The diversity of bacterial communities associated with M. laxissima increased significantly in aquaculture. Our work shows that it is important to monitor changes in bacterial communities when examining the feasibility of growing sponges in aquaculture systems because these communities may change. This could have implications for the health of sponges or for the production of bioactive compounds by sponges in cases where these compounds are produced by symbiotic bacteria rather than by the sponges themselves.     

Analysis of bacterial communities associated with spores of Gigaspora margarita and Gigaspora rosea

The spores of arbuscular mycorrhizal fungi (AMF) form a unique microhabitat that is suitable for the colonization by many species of bacteria. The aim of the current study was to analyze the bacterial communities associated with the surface of spores of the AMF species Gigaspora margarita MAFF 520054 and Gigaspora rosea JP1. The two AMF species were propagated with tobacco (Nicotiana tabacum) grown in a mixture of sand and soil. In another experiment, G. margarita was propagated with tobacco or alfalfa (Medicago sativa) grown in vermiculite or a mixture of sand and soil. The bacterial community composition of the new-formed spores and sand/soil substrate was analyzed using PCR of 16S rDNA fragments and denaturing gradient gel electrophoresis (DGGE). Clustering analysis revealed that the bacterial communities on the surface of G. margarita spores was different form that in the substrate or on the surface of the G. rosea spores, and both the host plant and the substrate could influence the composition of spore-associated bacterial populations of the G. margarita. Sequence analysis of the major DGGE bands of G. margarita spore samples revealed that most of the bacterial sequences were affiliated with the phyla Proteobacteria (Azospirillum, Azovibrio, Polyangium, Ramlibacter, Rubrivivax, Sphingomonas, and Rhizobium) and Actinobacteria (Streptomyces, Amycolatopsis, and Pseudonocardia).     

Use of horizontal subsurface flow constructed wetlands to treat reverse osmosis concentrate of rolling wastewater

According to the characteristics of the reverse osmosis concentrate (ROC) generated from iron and steel company, we used three sets of parallel horizontal subsurface flow (HSF) constructed wetlands (CWs) with different plants and substrate layouts to treat the high-salinity wastewater. The plant growth and removal efficiencies under saline condition were evaluated. The evaluation was based entirely on routinely collected water quality data and the physical and chemical characteristics of the plants (Phragmites australis, Typha latifolia, Iris wilsonii, and Scirpus planiculmis). The principal parameters of concern in the effluent were chemical oxygen demand (COD), total nitrogen (TN), and total phosphorus (TP). The results showed that the CWs were able to remove COD, TN, and TP from ROC. S. planiculmis was not suitable for the treatment of high-saline wastewater. The sequence of metals accumulated in CW plants was K>Ca>Na>Mg>Zn>Cu. More than 70% of metals were accumulated in the aboveground of P. australis. The CW filled with gravel and manganese ore and planted with P. australis and T. latifolia had the best performance of pollutant removal, with average removal of 49.96%, 39.45%, and 72.01% for COD, TN, and TP, respectively. The effluent water quality met the regulation in China. These results suggested that HSF CW planted with P. australis and T. latifolia can be applied for ROC pollutants removal.     

Spatial Variation of Phosphorous Retention Capacity in Subsurface Flow Constructed Wetlands: Effect of Wetland Type and Inflow Loading

For verification of spatial distribution of phosphorous retention capacity in constructed wetlands systems(CWs), two horizontal subsurface flow(HSSF) CWs and two vertical subsurface flow(VSSF) CWs, using sand as substrate and Typha latifolia as wetland plants, were constructed and put into use for synthetic wastewater treatment. Five months later, significant spatial variations of TP and inorganic phosphorus(Ca-P, Fe-P and Al-P) were observed, which were found to be greatly affected by CWs type and hydraulic loading. The results revealed that though spatial distribution of Fe-P and Al-P displayed a similar order of substrate content as "rhizosphere" > "near-rhizosphere" > "non-rhizosphere" and "inflow section" > "outflow section" regardless of types and loading, the distribution of Ca-P was positively correlated to that of Fe-P and Al-P in HSSF CWs, while negative correlation was shown in VSSF CWs. As a result, TP spatial distribution in HSSF CWs demonstrated a greater dissimilarity than that in VSSF CWs. For HSSF CWs with low hydraulic loading, the lowest TP content was found in non-rhizosphere substrate of outflow section, while the highest one was discovered in rhizonsphere substrate of inflow section. The values in 6 parts of areas ranged from 0.138 g·kg^-1 to 2.710 g·kg^-1, which also were from -33.5% to 1209% compared to the control value. On contrast, spatial difference of TP content in substrates of VSSF CWs was insignificant, with a variation ranging from 0.776 g·kg^-1 to 1.080 g·kg^-1, that was 275% to 421% higher than the control value. In addition, when hydraulic loading was increased, TP content in VSSF CWs sharply decreased, ranging from 0.210 g·kg^-1 to 0.634 g·kg^-1. Meanwhile, dissimilarity of TP spatial distribution in HSSF CWs was reduced, with TP content ranging from 0.258 g·kg^-1 to 2.237 g·kg^-1. The results suggested that P spatial distribution should be taken into account for CWs design and operation.     

Comparative study on removal of enteric pathogens from domestic wastewater using Typha latifolia and Cyperus rotundus along with different substrates

A comparative study was carried out to evaluate the efficiency of different substrate materials along with macrophytes Typha latifolia and Cyperus rotundus in treating domestic wastewater intended for reuse in agriculture. The study was conducted over a period of 6 months with different retention times, and observations were taken twice per month. One-way analysis of variance and Tukey's Honest Significant Difference (HSD) tests were used to determine statistical significant differences between experimental columns. Treatment with T. latifolia planted in sand and mix substrate with 4-day retention time remarkably reduced the concentration of all bacterial pathogens. Log reductions observed were approximately 5.01 and 4.82 for total coliform (TC), 4.46 and 3.93 for Escherichia coli, and 5.52 and 5.48 for Shigella, respectively. Moreover, these treatments were also efficient in completely removing fecal coliform (FC) and Salmonella.Maximum parasites were removed by the treatment having sand alone as a substrate containing C. rotundus, but the difference was not significant from those planted with T. latifolia in the same substrate. The results suggest that T. latifolia aids in bacterial pathogens removal, while C. rotundus aids in parasites removal. Thus, wastewater treatment through constructed wetland having mix plantation of these species along with sand can eliminate some of the major enteric pathogens.     

Mucosa-Associated Bacteria in the Human Gastrointestinal Tract Are Uniformly Distributed along the Colon and Differ from the Community Recovered from Feces

The human gastrointestinal (GI) tract harbors a complex community of bacterial cells in the mucosa, lumen, and feces. Since most attention has been focused on bacteria present in feces, knowledge about the mucosa-associated bacterial communities in different parts of the colon is limited. In this study, the bacterial communities in feces and biopsy samples from the ascending, transverse, and descending colons of 10 individuals were analyzed by using a 16S rRNA approach. Flow cytometric analysis indicated that 10⁵ to 10⁶ bacteria were present in the biopsy samples. To visualize the diversity of the predominant and the Lactobacillus group community, denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA gene amplicons was performed. DGGE analysis and similarity index comparisons demonstrated that the predominant mucosa-associated bacterial community was host specific and uniformly distributed along the colon but significantly different from the fecal community (P < 0.01). The Lactobacillus group-specific profiles were less complex than the profiles reflecting the predominant community. For 6 of the 10 individuals the community of Lactobacillus-like bacteria in the biopsy samples was similar to that in the feces. Amplicons having 99% sequence similarity to the 16S ribosomal DNA of Lactobacillus gasseri were detected in the biopsy samples of nine individuals. No significant differences were observed between healthy and diseased individuals. The observed host-specific DGGE profiles of the mucosa-associated bacterial community in the colon support the hypothesis that host-related factors are involved in the determination of the GI tract microbial community.     

Treatment of industrial wastewater with two-stage constructed wetlands planted with Typha latifolia and Phragmites australis

Industrial wastewater treatment comprises several processes to fulfill the discharge permits or to enable the reuse of wastewater. For tannery wastewater, constructed wetlands (CWs) may be an interesting treatment option. Two-stage series of horizontal subsurface flow CWs with Phragmites australis (UP series) and Typha latifolia (UT series) provided high removal of organics from tannery wastewater, up to 88% of biochemical oxygen demand (BOD₅) (from an inlet of 420 to 1000 mg L⁻¹) and 92% of chemical oxygen demand (COD) (from an inlet of 808 to 2449 mg L⁻¹), and of other contaminants, such as nitrogen, operating at hydraulic retention times of 2, 5 and 7 days. No significant (P < 0.05) differences in performance were found between both the series. Overall mass removals of up to 1294 kg COD ha⁻¹ d⁻¹ and 529 kg BOD₅ ha⁻¹ d⁻¹ were achieved for a loading ranging from 242 to 1925 kg COD ha⁻¹ d⁻¹ and from 126 to 900 kg BOD₅ ha⁻¹ d⁻¹. Plants were resilient to the conditions imposed, however P. australis exceeded T. latifolia in terms of propagation.     

Comparative studies to assess bacterial communities on the clover phylloplane using MLST, DGGE and T-RFLP

Denaturing gradient gel electrophoresis (DGGE) and terminal restriction fragment length polymorphism (T-RFLP) were used to characterise the changes that occurred in Bacillus cereus group strains present in the phylloplane of clover Trifolium hybridum over 4 months. These strains had previously been analysed by multiple locus sequence typing (MLST). DGGE displayed many equally intense bands which indicated many equally abundant ribotypes. The bacterial community composition was variable and the leaves sampled as little as a week apart were found to have some temporal variability, indicating that diverse phylloplane bacterial communities follow sequential patterns from time to time. The B. cereus group community clearly clustered into early, mid and late branches, possibly due to multiple successional sequences occurring during growing seasons. The functionally and phylogenetically diverse microbial communities appeared to exhibit predictable successional patterns over shorter time scales. DGGE analysis with the molecular marker rpoB gave better resolution than 16S rRNA amplicons. There were no strong similarities between the dendrograms produced by DGGE, MLST and T-RFLP and the clustering produced by the automated T-RFLP method was variable even between the three restriction enzymes used. The DGGE–MLST method emerged as a superior method to T-RFLP–MLST for rapid typing of bacterial communities.     

Study of microbial community structures in UASB sludge treating municipal wastewater by denaturing gradient gel electrophoresis of 16S rDNA

The structures of microbial communities in lab-scale upflow anaerobic sludge blanket (UASB) reactors for treating municipal wastewater with different ratios of COD soluble/COD total were studied using denaturing gradient gel electrophoresis (DGGE) of 16S rRNA genes.The microbial structure of the inoculum sludge obtained from a full-scale UASB reactor of treating potato processing wastewater was compared with the structures of sludges collected from three lab-scale UASB reactors after eight months feeding with raw municipal wastewater, with CEPS (chemically enhanced primary sedimentation) pretreated municipal wastewater, and with a synthetic municipal sewage, respectively. Computer-aided numerical analysis of the DGGE fingerprints showed that the bacterial community underwent major changes. The sludges for treating raw and CEPS pretreated wastewater had very similar bacterial and archaeal communities (82%and 96% similarity) but were different from that for treating the synthetic sewage. Hence, despite similar % COD in the particulate form in the synthetic and the real wastewater, the two wastewaters were selected for different microbial communities. Prominent DGGE bands of Bacteria and Archaea were purified and sequenced. The 16S rRNA gene sequences of the dominant archaeal bands found in the inoculum, and UASB sludge fed with raw sewage, CEPS pretreated wastewater, and synthetic sewage were closely associated with Methanosaeta concilii. In the UASB sludge fed with synthetic sewage, another dominant band associated with an uncultured archaeon 39-2 was found together with M. concilii.     

Endophytic bacteria in long-term in vitro cultivated “axenic” pineapple microplants revealed by PCR–DGGE

In vitro propagated plants are believed to be free of microbes. However, after 5 years of in vitro culture of pineapple plants, without evidence of microbial contamination, the use of culture-independent molecular approach [classifying heterogeneous nucleic acids amplified via universal and specific 16S rRNA gene by polymerase chain reaction (PCR)], and further analysis by denaturing gradient gel electrophoresis (DGGE) revealed endophytic bacteria in roots, young and mature leaves of such plants. The amplification of 16S rRNA gene (Bacteria domain) with the exclusion of the plant chloroplast DNA interference, confirmed the presence of bacterial DNA, from endophytic microorganisms within microplant tissues. PCR–DGGE analysis revealed clear differences on bacterial communities depending on plant organ. Group-specific DGGE analyses also indicated differences in the structures of Actinobacteria, Alphaproteobacteria and Betaproteobacteria communities in each part of plants. The results suggest the occurrence of a succession of bacterial communities colonizing actively the microplants organs. This study is the first report that brings together evidences that pineapple microplants, previously considered axenic, harbor an endophytic bacterial community encompassing members of Actinobacteria, Alphaproteobacteria and Betaproteobacteria group which is responsive to differences in organs due to plant development.     

Analysis of unculturable bacterial communities in tea orchard soils based on nested PCR-DGGE

The bacterial communities in the soils from tea orchards and their adjacent wasteland in Anhui Province, China were analysed by nested PCR-DGGE technique combined with sequencing. DGGE profiles revealed that the DGGE patterns of different soils were similar to each other and the most intensely bands appeared in all lanes. The bacterial genetic diversity index of tea orchard soils was lower than that of wasteland. For the tea orchard soils, Shannon’s diversity index decreased in the order: 45-year-old tea orchard >25-year-old tea orchard >7-year-old tea orchard >70-year-old tea orchard. The analysis of 16S rRNA gene sequences indicated that the fragments belong to Proteobacteria, Acidobacteria, TM7, Cyanobacteria and Firmicutes. A comprehensive analysis of the bacterial community structure in the tea orchard soils indicated the bacterial community was dominantly composed of Acidobacteria, followed by Proteobacteria (Gamma and Alpha), Firmicutes, Cyanobacteria and candidate division TM7. The RDA combined with UPGMA clustering analysis showed that the more similar the environmental variables were, the more similar the bacterial community structures in tea orchard soils were.     

Study of microbial community structures in UASB sludge treating municipal wastewater by denaturing gradient gel electrophoresis of 16S rDNA

The structures of microbial communities in lab-scale upflow anaerobic sludge blanket (UASB) reactors for treating municipal wastewater with different ratios of COD soluble/ COD total were studied using denaturing gradient gel electrophoresis (DGGE) of 16S rRNA genes. The microbial structure of the inoculum sludge obtained from a full-scale UASB reactor of treating potato processing wastewater was compared with the structures of sludges collected from three lab-scale UASB reactors after eight months feeding with raw municipal wastewater, with CEPS (chemically enhanced primary sedimentation) pretreated municipal wastewater, and with a synthetic municipal sewage, respectively. Computer-aided numerical analysis of the DGGE fingerprints showed that the bacterial community underwent major changes. The sludges for treating raw and CEPS pretreated wastewater had very similar bacterial and archaeal communities (82% and 96% similarity) but were different from that for treating the synthetic sewage. Hence, despite     

Constructed Wetlands for Tannery Wastewater Treatment in Portugal: Ten Years of Experience

Wastewaters from tannery industry are complex in composition and providing adequate treatment can be difficult. Constructed wetlands (CW) are regarded as an alternative treatment to the conventional biological systems, as a developing cost-effective and environmentally friendly phytoremediation technology. The present review compiles and integrates information on CWs technology for the needs of the tannery sector. The following issues arise as crucial for the implementation of such systems, namely i) an accurate wastewater characterization and an effective pretreatment before reaching the CW, ii) choosing the plants species better adapted to the imposed conditions, iii) substrate selection and iv) range of organic loadings applied. The examples practiced in Portugal give indication that horizontal subsurface flow systems, with expanded clay media, are a suitable option to be considered when dealing with high organic loading tannery wastewater (up to c.a. 3800 kgCODha^?1d^?1), being resilient to a wide range of hydraulic variations. Plants such as Phragmites and Typha have shown to be adequate for tannery wastewater depuration, with Arundo donax proving resilient to high salinity wastewaters. The flexibility of implementation allows the CW to be adapted to different sites with different configurations, being suitable as main secondary or tertiary treatment stage.     

Bacterial Population Changes in a Membrane Bioreactor for Graywater Treatment Monitored by Denaturing Gradient Gel Electrophoretic Analysis of 16S rRNA Gene Fragments

The bacterial population of a graywater treatment system was monitored over the course of 100 days, along with several wastewater biochemical parameters. The graywater treatment system employed an 1,800-liter membrane bioreactor (MBR) to process the waste, with essentially 100% recycling of the biomass. Graywater feed consisting of 10% galley water and 90% laundry water, selected to approximate the graywater composition on board U.S. Navy ships, was collected offsite. Five-day biological oxygen demand (BOD₅), oils and greases (O/G), nitrogen, and phosphorus were monitored in the feed and were found to vary greatly day to day. Changes in the bacterial population were monitored by PCR amplification of region 332 to 518 (Escherichia coli numbering) of the 16S rRNA gene and denaturing gradient gel electrophoresis (DGGE) analysis of the resultant PCR products. DGGE analysis indicated a diverse and unstable bacterial population throughout the 100-day period, with spikes in feed strength causing significant changes in community structure. Long-term similarity between the communities was 0 to 25%, depending on the method of analysis. In spite of the unstable bacterial population, the MBR system was able to meet effluent quality parameters approximately 90% of the time.     

Bacterial Community Structure in the Hyperarid Core of the Atacama Desert, Chile

Soils from the hyperarid Atacama Desert of northern Chile were sampled along an east-west elevational transect (23.75 to 24.70°S) through the driest sector to compare the relative structure of bacterial communities. Analysis of denaturing gradient gel electrophoresis (DGGE) profiles from each of the samples revealed that microbial communities from the extreme hyperarid core of the desert clustered separately from all of the remaining communities. Bands sequenced from DGGE profiles of two samples taken at a 22-month interval from this core region revealed the presence of similar populations dominated by bacteria from the Gemmatimonadetes and Planctomycetes phyla.     

Influence of Inorganic Nitrogen Management Regime on the Diversity of Nitrite-Oxidizing Bacteria in Agricultural Grassland Soils

To assess links between the diversity of nitrite-oxidizing bacteria (NOB) in agricultural grassland soils and inorganic N fertilizer management, NOB communities in fertilized and unfertilized soils were characterized by analysis of clone libraries and denaturing gradient gel electrophoresis (DGGE) of 16S rRNA gene fragments. Previously uncharacterized Nitrospira-like sequences were isolated from both long-term-fertilized and unfertilized soils, but DGGE migration patterns indicated the presence of additional sequence types in the fertilized soils. Detailed phylogenetic analysis of Nitrospira-like sequences suggests the existence of one newly described evolutionary group and of subclusters within previously described sublineages, potentially representing different ecotypes; the new group may represent a lineage of noncharacterized Nitrospira species. Clone libraries of Nitrobacter-like sequences generated from soils under different long-term N management regimes were dominated by sequences with high similarity to the rhizoplane isolate Nitrobacter sp. strain PJN1. However, the diversity of Nitrobacter communities did not differ significantly between the two soil types. This is the first cultivation-independent study of nitrite-oxidizing bacteria in soil demonstrating that nitrogen management practices influence the diversity of this bacterial functional group.