The Dynamic Response of Nitrogen Transformation to the Dissolved Oxygen Variations in the Simulated Biofilm Reactor

Lab-scale simulated biofilm reactors, including aerated reactors disturbed by short-term aeration interruption (AE-D) and non-aerated reactors disturbed by short-term aeration (AN-D), were established to study the stable-state (SS) formation and recovery after disturbance for nitrogen transformation in terms of dissolved oxygen (DO), removal efficiency (RE) of NH4+-N and NO3−-N and activity of key nitrogen-cycle functional genes amoA and nirS (RNA level abundance, per ball). SS formation and recovery of DO were completed in 0.56–7.75 h after transition between aeration (Ae) and aeration stop (As). In terms of pollutant REs, new temporary SS formation required 30.7–52.3 h after Ae and As interruptions, and seven-day Ae/As interruptions required 5.0% to 115.5% longer recovery times compared to one-day interruptions in AE-D and AN-D systems. According to amoA activity, 60.8 h were required in AE-D systems to establish new temporary SS after As interruptions, and RNA amoA copies (copy number/microliter) decreased 88.5%, while 287.2 h were required in AN-D systems, and RNA amoA copies (copy number/microliter) increased 36.4 times. For nirS activity, 75.2–85.8 h were required to establish new SSs after Ae and As interruptions. The results suggested that new temporary SS formation and recovery in terms of DO, pollutant REs and amoA and nirS gene activities could be modelled by logistic functions. It is concluded that temporary SS formation and recovery after Ae and As interruptions occurred at asynchronous rates in terms of DO, pollutant REs and amoA and nirS gene activities. Because of DO fluctuations, the quantitative relationship between gene activity and pollutant RE remains a challenge.

Spatial and Seasonal Variations in the Abundance of Nitrogen-Transforming Genes and the Microbial Community Structure in Freshwater Lakes with Different Trophic Statuses

Identifying nitrogen-transforming genes and the microbial community in the lacustrine sedimentary environment is critical for revealing nitrogen cycle processes in eutrophic lakes. In this study, we examined the diversity and abundance of ammonia-oxidizing bacteria (AOB), ammonia-oxidizing archaea (AOA), denitrifying bacteria (DNB), and anammox bacteria (AAOB) in different trophic status regions of Lake Taihu using the amoA, Arch-amoA, nirS, and hzo genes as functional markers. Quantitative Polymerase Chain Reaction (qPCR) results indicated that the abundance of the nirS gene was the highest, while the amoA gene had the lowest abundance in all regions. Except for the primary inflow area of Lake Taihu, Arch-amoA gene abundance was higher than the hzo gene in three lake bays, and the abundance of the nirS gene increased with decreasing trophic status. The opposite pattern was observed for the amoA, Arch-amoA, and hzo genes. Phylogenetic analyses showed that the predominant AOB and AOA were Nitrosomonas and Nitrosopumilus maritimus, respectively, and the proportion of Nitrosomonas in the eutrophic region (87.9%) was higher than that in the mesotrophic region (71.1%). Brocadia and Anammoxoglobus were the two predominant AAOB in Lake Taihu. Five novel unknown phylotypes of AAOB were observed, and Cluster AAOB-B was only observed in the inflow area with a proportion of 32%. In the DNB community, Flavobacterium occurred at a higher proportion (22.6–38.2%) in all regions, the proportion of Arthrobacter in the mesotrophic region (3.6%) was significantly lower than that in the eutrophic region (15.6%), and the proportions of Cluster DNB-E in the inflow area (24.5%) was significantly higher than that in the lake bay (7.3%). The canonical correspondence analysis demonstrated that the substrate concentration in sedimentary environments, such as NOx--N in the sediment, NH4+-N in the pore water, and the total organic matter, were the key factors that determined the nitrogen-transforming microbial community. However, the temperature was also a predominant factor affecting the AOA and AAOB communities.

Relationships between nitrogen removal processes and functional microorganisms in the rhizosphere soil in a horizontal surface flow constructed wetland

Plant species could significantly affect the nitrogen removal processes mediated by microorganisms in constructed wetlands. However, the links between nitrogen removal processes in the rhizosphere and the related functional microorganisms in a horizontal surface flow constructed wetland in winter remain poorly understood. In this study we collected 24 rhizosphere soils from Typha orientalis and Phragmites australis to evaluate potential nitrogen removal activities, namely the potential nitrification rate (PNR) and denitrification enzyme activity (DEA), and their relationship with functional genes (i.e. nitrate reductase, nirS, and ammonia mono-oxygenase, amoA, of ammonia-oxidising archaea, AOA, and ammonia-oxidising bacteria, AOB) in denitrifiers and nitrifiers in winter. DEA and PNR were significantly higher in the rhizosphere soil of T. orientalis than P. australis, which was due to the higher abundance of nitrifiers and denitrifiers in the rhizosphere of T. orientalis. AOB were the major predictor of PNR in rhizosphere soil of T. orientalis, whereas AOA were more important for P. australis. In addition, denitrifiers containing the nirS gene were found to be the main drivers of DEA, and AOA and AOB also contributed to the denitrification process in the rhizosphere soil of both plants. Furthermore, the abundance of nitrifiers was significantly affected by the C:N ratio, soil organic matter and moisture, whereas the abundance of denitrifiers was affected by soil moisture and pH.

Comparison of the Abundance and Community Structure of N-Cycling Bacteria in Paddy Rhizosphere Soil under Different Rice Cultivation Patterns

Eco-agricultural systems aim to reduce the use of chemical fertilizers in order to improve sustainable production and maintain a healthy ecosystem. The aim of this study was to explore the effects of rice-frog farming on the bacterial community and N-cycling microbes in paddy rhizosphere soil. This experiment involved three rice cultivation patterns: Conventionally cultivated rice (CR), green rice-frog farming (GR), and organic rice-frog farming (OR). The rice yield, paddy soil enzyme activities, physicochemical variables and bacterial and N-cycling bacterial abundances were quantitatively analyzed. Rice-frog cultivations significantly increased soil protease, nitrate and reductase activity. Additionally, the nirS gene copy number and the relative abundance of denitrifying bacteria also increased, however urease activity and the relative abundance of nitrifying bacteria significantly decreased. The bacterial community richness and diversity of OR soil was significantly higher than that of the GR or CR soil. Nitrogen use efficiency (NUE) of GR was highest. The N-cycling bacterial community was positively correlated with the total carbon (TC), total nitrogren (TN) and carbon to nitrogen (C:N) ratio. The present work strengthens our current understanding of the soil bacterial community structure and its functions under rice-frog farming. The present work also provides certain theoretical support for the selection of rational rice cultivation patterns.

Characterization and activity of endophytic bacteria from ‘Prata Anã’ banana crop (Musa sp., AAB)

ABSTRACT The objective of this study was to characterize banana tree endophytic bacteria at genus and species level and to determine the metabolic reactions associated with the nitrogen transformations. The identification at genus and species levels was performed using the partial sequencing of the rDNA 16S region. The assimbyotic nitrogen fixation, the reduction of nitrate and the production of urease were in vitro evaluated. The DNA of the bacterial isolates was also amplified to verify the presence of the nifH, nirK and nirS regions. Biochemical tests were performed in a complete randomized design; the treatments consisted of 39 bacterial isolates with three replications. Sequence analysis enabled the identification of four genera: Bacillus, Rhizobium, Klebsiella and Enterobacter. The Bacillus genus occurred more frequently, nine species were identified. By evaluating the results of biochemical tests, it was observed that three isolates showed multiple abilities: growth in NFb medium, nitrate reduction and production of urease. The isolates belong to the genus Bacillus and of the species subtilis, thuringienses and amyloliquefaciens. Approximately 12.5% of the isolates amplified the region corresponding to the nifH gene, 7.5% amplified gene nirK and 3.9% amplified the nirS gene. Endophytic bacteria evaluated in the present study showed in vitro activity for urease, nitrate reductase enzymes, however, relevant nitrogenase activity was not observed.

Distinct distribution patterns of proteobacterial nirK- and nirS-type denitrifiers in the Yellow River estuary, China

Denitrification is considered to be the critical process in removing reactive nitrogen in estuarine ecosystems. In the present study, the abundance, diversity, and community structure of nirK- and nirS-type denitrifiers were compared in sediments from the Yellow River estuary. Quantitative polymerase chain reaction showed that the 2 types of denitrifiers exhibited different distribution patterns among the samples, indicating their distinct habitat preference. Phylogenetic analysis revealed that most of the sequences from clusters I, III, IV, and V for nirK-type denitrifiers were dominant and were distributed at sites where dissolved oxygen (DO) was lower, and the sequences in the other clusters were dominant at sites with higher DO. However, there was no spatially heterogeneous distribution for the nirS-type denitrifier community. Canonical correlation analysis and correlation analysis demonstrated that the community structure of nirK was more responsive to environmental factors than was that of nirS. Inversely, the abundance and α-diversity targeting nirS gene could be more easily influenced by environmental parameters. These findings can extend our current knowledge about the distribution patterns of denitrifying bacteria and provide a basic theoretical reference for the dynamics of denitrifying communities in estuarine ecosystem of China.