Characterization of functional microbial community in a membrane-aerated biofilm reactor operated for completely autotrophic nitrogen removal

The 16S rDNA-based molecular technique was applied to investigate the functional microbial community of a membrane-aerated biofilm (MAB) that was used for completely autotrophic nitrogen removal over nitrite (CANON). The relationships among two kinds of key bacteria responsible for CANON: aerobic ammonia-oxidizing bacteria (AOB) and Anammox bacteria, and their possible distributions in the MAB were discussed based on the microbial community analysis. FISH analysis showed the existence of two visible active layers in experimental MAB. One is the partial nitrifying layer located in the region of oxygen-rich membrane-biofilm interface, dominated by NSO190-positive AOB. The other is the Anammox active layer located in the region of anoxic liquid-biofilm interface, dominated by PLA46 and AMX820-positive Anammox microorganisms. As a result of this study, the AOB as well as Anammox bacteria were present and active in experimental MABR, and the cooperation between AOB and Anammox bacteria was considered to be responsible for CANON.     

Microscale heterogeneity of the soil nitrogen cycling microbial functional structure and potential metabolism

Soil aggregates, with complex spatial and nutritional heterogeneity, are clearly important for regulating microbial community ecology and biogeochemistry in soils. However, how the taxonomic composition and functional attributes of N-cycling-microbes within different soil particle-size fractions under a long-term fertilization treatment remains largely unknown. Here, we examined the composition and metabolic potential for urease activity, nitrification, N₂O production and reduction of the microbial communities attached to different sized soil particles (2000–250, 250–53 and <53 μm) using a functional gene microarray (GeoChip) and functional assays. We found that urease activity and nitrification were higher in <53 μm fractions, whereas N₂O production and reduction rates were greater in 2000–250 and 250–53 μm across different fertilizer regimes. The abundance of key N-cycling genes involved in anammox, ammonification, assimilatory and dissimilatory N reduction, denitrification, nitrification and N₂-fixation detected by GeoChip increased as soil aggregate size decreased; and the particular key genes abundance (e.g., ureC, amoA, narG, nirS/K) and their corresponding activity were uncoupled. Aggregate fraction exerted significant impacts on N-cycling microbial taxonomic composition, which was significantly shaped by soil nutrition. Taken together, these findings indicate the important roles of soil aggregates in differentiating N-cycling metabolic potential and taxonomic composition, and provide empirical evidence that nitrogen metabolism potential and community are uncoupled due to aggregate heterogeneity.     

土壤微生物生物量氮及其在氮素循环中作用

简述了土壤微生物生物量氮的含量及其影响因素,阐述了其在土壤氮素循环中的重要作用,着重讨论了其与可矿化氮、矿质氮、有机氮和固定态铵之间的关系,指出土壤微生物生物量氮与供氮因子间的关系在氮素循环研究中有非常重要的作用,可为调控土壤氮素的供应状况,减少氮素损失,提高氮肥利用率提供科学依据,并提出了需要深入研究的问题。     

Abundances and potential activities of nitrogen cycling microbial communities along a chronosequence of a glacier forefield

Glacier forefields are ideal ecosystems to study the development of nutrient cycles as well as single turnover processes during soil development. In this study, we examined the ecology of the microbial nitrogen (N) cycle in bulk soil samples from a chronosequence of the Damma glacier, Switzerland. Major processes of the N cycle were reconstructed on the genetic as well as the potential enzyme activity level at sites of the chronosequence that have been ice-free for 10, 50, 70, 120 and 2000 years. In our study, we focused on N fixation, mineralization (chitinolysis and proteolysis), nitrification and denitrification. Our results suggest that mineralization, mainly the decomposition of deposited organic material, was the main driver for N turnover in initial soils, that is, ice-free for 10 years. Transient soils being ice-free for 50 and 70 years were characterized by a high abundance of N fixing microorganisms. In developed soils, ice-free for 120 and 2000 years, significant rates of nitrification and denitrification were measured. Surprisingly, copy numbers of the respective functional genes encoding the corresponding enzymes were already high in the initial phase of soil development. This clearly indicates that the genetic potential is not the driver for certain functional traits in the initial phase of soil formation but rather a well-balanced expression of the respective genes coding for selected functions.     

Performance of a membrane biofilm reactor for denitrification with methane

In this study, a membrane biofilm reactor was investigated for aerobic methane oxidation coupled indirectly to denitrification, a process potentially useful for denitrification of nitrate-contaminated waters and wastewaters using methane as external electron donor. Methane and oxygen were supplied from the interior of a silicone tube to a biofilm growing on its surface. We found that the membrane biofilm reactor was to some extent self-regulating in the supply of methane and oxygen. Although the intramembrane partial pressures of methane and oxygen were varied, the oxygen-to-methane ratio penetrating the membrane tended towards 1.68. Both nitrate removal rate and dissolved organic carbon (DOC) production rate appeared to be positively correlated with intramembrane methane pressure. Based on measured nitrate removal rates, DOC production rates, and nitrate removal efficiency, the possibility of using this method for treatment of a hypothetical wastewater was evaluated.     

Nitrogen removal performance and microbial community structure dynamics response to carbon nitrogen ratio in a compact suspended carrier biofilm reactor

A compact suspended carrier biofilm reactor (SCBR) was developed for simultaneous nitrification and denitrification (SND) in a single reactor and the performance of nutrient removal was investigated. Microbial community structure response to different ratio of carbon to nitrogen (C/N) was determined by denaturing gel gradient electrophoresis (DGGE) profiles of 16S rDNA V3 region and amoA gene amplifications. In addition, the population dynamics of ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) were estimated by fluorescence in situ hybridization (FISH) with 16S rDNA-targeted oligonucleotide probes. Results showed that the compact SCBR was efficient in nutrient removal with COD_Cr removal efficiency over 90% and SND efficiency (E_SND) about 83.3%. The diversity of microbial community structure was positively correlated with C/N ratio, while the three communities of amoA gene were relativity homogenous. The population of nitrifiers was in inverse proportions to C/N ratio with the average fraction of AOB and NOB to all bacteria 5.4, 4.8, 3.1% and 4.6, 3.5, 2.7% respectively as C/N ratio changing from 3:1, 5:1 to 10:1. Therefore we could reach a conclusion that the compact SCBR was practical to treat municipal wastewater and the shift of microbial community monitored by molecular technologies could offer guidance to the process optimization in engineering.     

Kinetics of microbial bromate reduction in a hydrogen-oxidizing, denitrifying biofilm reactor

Bromate (BrO(3)(-)) is an oxidized contaminant produced from bromide (Br(-)) during ozonation and advanced oxidation of drinking water. Previous research shows that denitrifying bioreactors can reduce bromate to innocuous bromide. We studied a hydrogen-based, denitrifying membrane-biofilm reactor (MBfR) for bromate reduction, and report the first kinetics for a hydrogen-based bromate reduction process. A mixed-culture MBfR reduced up to 1,500 microg/L bromate to below 10 microg/L with a 50-min hydraulic residence time. Kinetics were determined using short-term tests on a completely mixed MBfR at steady state with an influent of 5 mg N/L nitrate plus 100 microg/L bromate. Short-term tests examined the impact of pH, nitrite, nitrate, and bromate on bromate reduction rates in the MBfR. Kinetic parameters for the process were estimated based on the short-term bromate tests. The q(max) for bromate reduction was 0.12 mg BrO(3)(-) x mg(x)(-1) x day(-1), and the K was 1.2 mg BrO(3)(-)/L. This q(max) is 2-3 times higher than reported for heterotrophic enrichments, and the K is the first reported in the literature. Nitrite and nitrate partially inhibited bromate reduction, with nitrite exerting a stronger inhibitory effect. Bromate was self-inhibitory at concentrations above 15 mg/L, but up to 50 mg/L of bromate had no inhibitory effect on denitrification. The optimum pH was approximately 7. We also examined the performance of an MBfR containing pure culture of the denitrifying bacterium Ralstonia eutropha. Under conditions similar to the mixed-culture tests, no bromate reduction was detected, showing that not all denitrifying bacteria are active in bromate reduction. Our results suggest the presence of specialized, dissimilatory bromate-reducing bacteria in the mixed-culture MBfR.     

High-strength wastewater treatment using microbial biofilm reactor: a critical review

World Journal of Microbiology and Biotechnology - Natural products extracted from plants are an alternative method for controlling postharvest citrus blue mold, caused by Penicillium italicum (P....     

Phenol and sulfide oxidation in a denitrifying biofilm reactor and its microbial community analysis

A microbial consortium attached onto a polyethylene support was used to evaluate the simultaneous oxidation of sulfide and phenol by denitrification. The phenol, sulfide and nitrate loading rates applied to an inverse fluidized bed reactor were up to 168 mg phenol–C/(l d), 37 mg S^2?/(l d) and 168 mg NO₃^?–N/(l d), respectively. Under steady state operation the consumption efficiencies of phenol, sulfide and nitrate were 100%. The N₂ yield (g N₂/g NO₃^?–N) was 0.89. The phenol was mineralized resulting in a yield of 0.82 g bicarbonate–C/g phenol–C and sulfide was completely oxidized to sulfate with a yield of 0.99 g SO₄^2?–S/g S^2?. 16S rRNA gene-based microbial community analysis of the denitrifying biofilm showed the presence of Thauera aromatica, Thiobacillus denitrificans, Thiobacillus sajanensis and Thiobacillus sp. This is the first work reporting the simultaneous oxidation of sulfide and phenol in a denitrifying biofilm reactor.     

Leaf litter diversity alters microbial activity,microbial abundances,and nutrient cycling in a subtropical forest ecosystem

Human activities affect both tree species composition and diversity in forested ecosystems. This in turn alters the species diversity of plant litter and litter quality, which may have cascading effects on soil microbial communities and their functions for decomposition and nutrient cycling. We tested microbial responses to litter species diversity in a leaf litter decomposition experiment including monocultures, 2-, and 4-species mixtures in the subtropical climate zone of southeastern China. Soil microbial community composition was assessed by lipid analysis, and microbial functions were measured using extracellular enzyme activity and gross rates of nitrogen mineralization. We observed a positive relationship between litter species diversity and abundances of mycorrhizal fungi and actinomycetes. Alternatively, enzyme activities involved in carbon and phosphorus acquisition, and enzyme indices of relative carbon limitation, were higher only in the 4-species mixtures. This suggests that the minimum basal substrate level for enzyme production was reached, or that limitation was higher, at the highest diversity level only. Responses to litter diversity also changed over time, where phosphatase responses to litter diversity were strongest early in decomposition and the indices of carbon limitation relative to other nutrients showed stronger responses later in decomposition. Enzyme activities were related to lipid biomarker data and the mass of litter remaining at the third time point, but relationships between enzyme activity and the mass of litter remaining were not consistent across other time points. We conclude that litter species richness will likely only reduce microbial functions at key intervals of diversity loss while microbial growth is more sensitive to incremental diversity loss, with no clear relationships between them or to ecosystem functions. The observed litter diversity effects on soil microbial biomass and enzyme activity indicate interactions of aboveground and belowground communities, and together with environmental conditions they are important for maintaining ecosystem functions.     

Loss in microbial diversity affects nitrogen cycling in soil

Microbial communities have a central role in ecosystem processes by driving the Earth''s biogeochemical cycles. However, the importance of microbial diversity for ecosystem functioning is still debated. Here, we experimentally manipulated the soil microbial community using a dilution approach to analyze the functional consequences of diversity loss. A trait-centered approach was embraced using the denitrifiers as model guild due to their role in nitrogen cycling, a major ecosystem service. How various diversity metrics related to richness, eveness and phylogenetic diversity of the soil denitrifier community were affected by the removal experiment was assessed by 454 sequencing. As expected, the diversity metrics indicated a decrease in diversity in the 1/10³ and 1/10⁵ dilution treatments compared with the undiluted one. However, the extent of dilution and the corresponding reduction in diversity were not commensurate, as a dilution of five orders of magnitude resulted in a 75% decrease in estimated richness. This reduction in denitrifier diversity resulted in a significantly lower potential denitrification activity in soil of up to 4–5 folds. Addition of wheat residues significantly increased differences in potential denitrification between diversity levels, indicating that the resource level can influence the shape of the microbial diversity–functioning relationship. This study shows that microbial diversity loss can alter terrestrial ecosystem processes, which suggests that the importance of functional redundancy in soil microbial communities has been overstated.     

Organic carbon and nitrogen removal from a strong wastewater using a denitrifying suspended growth reactor and a horizontal-flow biofilm reactor

Recirculation of fully nitrified effluent from a laboratory horizontal-flow biofilm reactor (HFBR) to a mixed pre-denitrification reactor (DR) was used to remove organic carbon and nitrogen from synthetic dairy wastewater. Three recirculation ratios of 2, 4, and 6 were examined in this study and the average filtered chemical oxygen demand (CODf) and total nitrogen (TN) removals were up to 97.4% and 85.5%, respectively, at 11 degrees C. In the DR, the nitrate nitrogen removal efficiencies and rates were 86-96% and 22-34 g N/m3 d. In the HFBR, the ammonium nitrogen removal rates were 293-337 mg N/m2 d.     

Backwash intensity and frequency impact the microbial community structure and function in a fixed-bed biofilm reactor

Linkages among bioreactor operation and performance and microbial community structure were investigated for a fixed-bed biofilm system designed to remove perchlorate from drinking water. Perchlorate removal was monitored to evaluate reactor performance during and after the frequency and intensity of the backwash procedure were changed, while the microbial community structure was studied using clone libraries and quantitative PCR targeting the 16S rRNA gene. When backwash frequency was increased from once per month to once per day, perchlorate removal initially deteriorated and then recovered, and the relative abundance of perchlorate-reducing bacteria (PRB) initially increased and then decreased. This apparent discrepancy suggested that bacterial populations other than PRB played an indirect role in perchlorate removal, likely by consuming dissolved oxygen, a competing electron acceptor. When backwash intensity was increased, the reactor gradually lost its ability to remove perchlorate, and concurrently the relative abundance of PRB decreased. The results indicated that changes in reactor operation had a profound impact on reactor performance through altering the microbial community structure. Backwashing is an important yet poorly characterized procedure when operating fixed-bed biofilm reactors. Compared to backwash intensity, changes in backwash frequency exerted less disturbance on the microbial community in the current study. If this finding can be confirmed in future work, backwash frequency may serve as the primary parameter when optimizing backwash procedures.     

Effect of monospecific and mixed Cunninghamia lanceolata plantations on microbial community and two functional genes involved in nitrogen cycling

Conversion of native broadleaf forest (NF) and introduction of broadleaf species into monospecific Cunninghamia lanceolata plantations are silvicultural practices driven by the increasing demand for timber production. This study was conducted to evaluate the impacts of successive planting of C. lanceolata and mixed plantations (C. lanceolata-Michelia macclurei, CFM; C. lanceolata-Alnus cremastogyne, CFA; C. lanceolata-Kalopanax septemlobus, CFK) on microbial community diversity. Microbial biomass (MBC) was assessed using chloroform fumigation-extraction. Using denaturing gradient gel electrophoresis (DGGE), we examined the biodiversity within eubacterial (16S rRNA gene) and fungal (28S rRNA gene) species and two genes involved in N cycling: nifH and amoA. Microbial community diversities and microbial biomass decreased as NF was substituted by successive plantings of C. lanceolata plantations, whereas the trend reversed after introducing the broadleaf, M. macclurei, into pure C. lanceolata plantations. A strong positive correlation between MBC changes and total organic C (TOC), total organic N (TON), available N and extractable C (C_ext) were seen, which suggests that MBC was tightly coupled with the content of soil organic matter. The Shannon index showed that bacterial diversity and two functional genes (nifH and amoA) diversities associated with monospecific C. lanceolata plantations were lower than that of NF or mixed C. lanceolata plantations, such as CFM and CFA, whereas the opposite was seen for fungal diversity. Bacterial diversity was positively correlated with pH, TOC, TON, C_ext and NH ₄ ⁺ -N; while fungal diversity was positively correlated with C/N ratio and negatively correlated with pH. Both nitrogen fixing and ammonia oxidizing bacterial diversities were positively correlated with pH. Thus, soil pH was not only significantly positively correlated with bacterial diversity (r?=?0.502, P?<?0.05), nifH gene diversity (r?=?0.564, P?<?0.01) and amoA gene diversity (r?=?0.659, P?<?0.001), but also negatively correlated with fungal diversity (r?=?? 0.505, P?<?0.05), which seemed to be responsible for the discrimination of the soil microbial communities among these plantations. These findings suggest that different silvicultural practices have significant impacts on the soil microbial community through influences on soil chemical properties.     

Microwave radiation and reactor design influence microbial communities during methane fermentation

The effect of reactor design and method of heating on the efficiency of methane fermentation and composition of microbial communities, especially methanogenic Archaea, were determined. The research was carried out using submerge- and trickling-bed reactors fed with wastewater and the heat supply into the reactors included a convection heating method and microwave radiation. The polymerase chain reaction-denaturing gradient gel electrophoresis and relative real-time PCR were used in order to assess the biofilm communities. The best fermentation results and the highest abundance of methanogenic Archaea in biomass were observed in microwave heated trickling-bed reactors. The research proved that in reactors of identical design, the application of microwaves enabled a higher fermentation efficiency to be obtained and simultaneously increased the diversity of methanogenic Archaea communities that favors process stability. All the identified sequences of Archaea belonged to Methanosarcina sp., suggesting that species from this genera are susceptible to non-thermal effects of microwaves. There were no effects from microwaves on the bacterial communities in both types of reactors, however, the bacterial species composition varied in the reactors of different design.     

Utilisation of a packed-bed biofilm reactor for the determination of the potential of biofilm accumulation in water systems

An experimental system has been developed that allows the monitoring of biofilm development on supports exposed to water of different characteristics. The system consists of a series of packed-bed reactors filled with glass beads, and by periodically removing biofilm attached to these beads for off-line analyses this provides a means for monitoring biofilm development. Despite its reduced dimensions (6.9 cm long and 1.58 cm in diameter), the experimental system used has a sampling surface of 90.3 cm2 (including only the surface of the glass beads). This allows reproducible and representative samples to be taken from different water systems, providing a reliable and economic method for evaluating in situ the formation of biofilms from different environments. The set-up of the entire experimental system was constructed to meet the demands of field experiments in a well-defined hydrodynamic environment and to allow easy removal of samples for biomass quantification and microscopic observation. Data obtained using this device can be used as an indicator of the risk of biofilm formation in different water systems. This indicator, "the biofilm accumulation potential", represents an effective and representative tool for the monitoring of biofilm development in an integrated antifouling strategy, in order to help keep biofouling, scaling and microbial risks under control. According to the experiments with the packed-bed reactors used with a high flow regime, the ratio TCN/HPC could provide an indication of the state of the biofilm, and lower ratios could indicate a higher biofilm accumulation potential.     

Utilisation of a packed-bed biofilm reactor for the determination of the potential of biofilm accumulation in water systems

Abstract

An experimental system has been developed that allows the monitoring of biofilm development on supports exposed to water of different characteristics. The system consists of a series of packed-bed reactors filled with glass beads, and by periodically removing biofilm attached to these beads for off-line analyses this provides a means for monitoring biofilm development. Despite its reduced dimensions (6.9 cm long and 1.58 cm in diameter), the experimental system used has a sampling surface of 90.3 cm² (including only the surface of the glass beads). This allows reproducible and representative samples to be taken from different water systems, providing a reliable and economic method for evaluating in situ the formation of biofilms from different environments. The set-up of the entire experimental system was constructed to meet the demands of field experiments in a well-defined hydrodynamic environment and to allow easy removal of samples for biomass quantification and microscopic observation. Data obtained using this device can be used as an indicator of the risk of biofilm formation in different water systems. This indicator, “the biofilm accumulation potential”, represents an effective and representative tool for the monitoring of biofilm development in an integrated antifouling strategy, in order to help keep biofouling, scaling and microbial risks under control. According to the experiments with the packed-bed reactors used with a high flow regime, the ratio TCN/HPC could provide an indication of the state of the biofilm, and lower ratios could indicate a higher biofilm accumulation potential.     

Kinetics of anaerobic methane oxidation coupled to denitrification in the membrane biofilm reactor

Anaerobic oxidation of methane coupled to denitrification (AOM-D) in a membrane biofilm reactor (MBfR), a platform used for efficiently coupling gas delivery and biofilm development, has attracted attention in recent years due to the low cost and high availability of methane. However, experimental studies have shown that the nitrate-removal flux in the CH₄-based MBfR (<1.0 g N/m²-day) is about one order of magnitude smaller than that in the H₂-based MBfR (1.1–6.7 g N/m²-day). A one-dimensional multispecies biofilm model predicts that the nitrate-removal flux in the CH₄-based MBfR is limited to <1.7 g N/m²-day, consistent with the experimental studies reported in the literature. The model also determines the two major limiting factors for the nitrate-removal flux: The methane half-maximum-rate concentration (K₂) and the specific maximum methane utilization rate of the AOM-D syntrophic consortium (k_max2), with k_max2 being more important. Model simulations show that increasing k_max2 to >3 g chemical oxygen demand (COD)/g cell-day (from its current 1.8 g COD/g cell-day) and developing a new membrane with doubled methane-delivery capacity (D_m) could bring the nitrate-removal flux to ≥4.0 g N/m²-day, which is close to the nitrate-removal flux for the H₂-based MBfR. Further increase of the maximum nitrate-removal flux can be achieved when D_m and k_max2 increase together.