A denitrifying microbial consortium was enriched in an anoxically operated, methanol-fed sequencing batch reactor (SBR) fed with a mineral salts medium containing methanol as the sole carbon source and nitrate as the electron acceptor. The SBR was inoculated with sludge from a biological nutrient removal activated sludge plant exhibiting good denitrification. The SBR denitrification rate improved from less than 0.02 mg of NO 3 ؊ -N mg of mixed-liquor volatile suspended solids (MLVSS)؊1 h ؊1 to a steady-state value of 0.06 mg of NO 3 ؊ -N mg of MLVSS ؊1 h ؊1 over a 7-month operational period. At this time, the enriched microbial community was subjected to stable-isotope probing (SIP) with 14 C]methanol uptake in the enriched biomass. The well-known denitrification lag period in the methanol-fed SBR was shown to coincide with a lag phase in growth of the DEN67-targeted denitrifying population. We conclude that Methylophilales bacteria are the dominant denitrifiers in our SBR system and likely are important denitrifiers in full-scale methanol-fed denitrifying sludges.
The acetate-utilizing microbial consortium in a full-scale activated sludge process was investigated without prior enrichment using stable isotope probing (SIP). [13 C]acetate was used in SIP to label the DNA of the denitrifiers. The [ 13 C]DNA fraction that was extracted was subjected to a full-cycle rRNA analysis. The dominant 16S rRNA gene phylotypes in the 13 C library were closely related to the bacterial families Comamonadaceae and Rhodocyclaceae in the class Betaproteobacteria. Seven oligonucleotide probes for use in fluorescent in situ hybridization (FISH) were designed to specifically target these clones. Application of these probes to the sludge of a continuously fed denitrifying sequencing batch reactor (CFDSBR) operated for 16 days revealed that there was a significant positive correlation between the CFDSBR denitrification rate and the relative abundance of all probe-targeted bacteria in the CFDSBR community. FISH-microautoradiography demonstrated that the DEN581 and DEN124 probe-targeted cells that dominated the CFDSBR were capable of taking up [14 C]acetate under anoxic conditions. Initially, DEN444 and DEN1454 probe-targeted bacteria also dominated the CFDSBR biomass, but eventually DEN581 and DEN124 probe-targeted bacteria were the dominant bacterial groups. All probe-targeted bacteria assessed in this study were denitrifiers capable of utilizing acetate as a source of carbon. The rapid increase in the number of organisms positively correlated with the immediate increase in denitrification rates observed by plant operators when acetate is used as an external source of carbon to enhance denitrification. We suggest that the impact of bacteria on activated sludge subjected to intermittent acetate supplementation should be assessed prior to the widespread use of acetate in the wastewater industry to enhance denitrification.Nonpolluted natural bodies of water, such as freshwater streams and coastal marine water, have been found to have limiting concentrations of both dissolved inorganic nitrogen and dissolved inorganic phosphorus (11). According to limiting-nutrient concepts, the growth of phytoplankton, macroalgae, seagrass, etc. is limited in these aquatic environments mainly due to the low availability of dissolved inorganic nitrogen and dissolved inorganic phosphorus. Dennison and Abal (11) demonstrated that stimulation of the growth of these organisms and a subsequent imbalance in the aquatic ecosystems is inevitable when dissolved inorganic nitrogen is not limiting in these bodies of water. Therefore, the control of nitrogen in bodies of water has been highlighted as an important environmental activity, and stringent standards for total nitrogen levels have been imposed for effluents released from wastewater treatment plants.Biological nitrogen removal from wastewater is a two-step, sequential process. The first step is nitrification, an aerobic process in which ammonia is oxidized to nitrate (27). The second step is denitrification to remove soluble nitrate and nitrite from the wastewater (39)...
Although health risk due to discoloured water is minimal, such water continues to be the source of one of the major complaints received by most water utilities in Australia. Elevated levels of iron (Fe) and/or manganese (Mn) in bulk water are associated with discoloured water incidents. The accumulation of these two elements in distribution systems is believed to be one of the main causes for such elevated levels. An investigation into the contribution of pipe wall biofilms towards Fe and Mn deposition, and discoloured water events is reported in this study. Eight laboratory-scale reactors were operated to test four different conditions in duplicate. Four reactors were exposed to low Fe (0.05 mg l(-1)) and Mn (0.02 mg l(-1)) concentrations and the remaining four were exposed to a higher (0.3 and 0.4 mg l(-1) for Fe and Mn, respectively) concentration. Two of the four reactors which received low and high Fe and Mn concentrations were chlorinated (3.0 mg l(-1) of chlorine). The biological activity (measured in terms of ATP) on the glass rings in these reactors was very low (∼1.5 ng cm(-2) ring). Higher concentrations of Fe and Mn in bulk water and active biofilms resulted in increased deposition of Fe and Mn on the glass rings. Moreover, with an increase in biological activity, an increase in Fe and Mn deposition was observed. The observations in the laboratory-scale experiments were in line with the results of field observations that were carried out using biofilm monitors. The field data additionally demonstrated the effect of seasons, where increased biofilm activities observed on pipe wall biofilms during late summer and early autumn were found to be associated with increased deposition of Fe and Mn. In contrast, during the cooler months, biofilm activities were a magnitude lower and the deposited metal concentrations were also significantly less (ie a drop of 68% for Fe and 86% for Mn). Based on the laboratory-scale investigations, detachment of pipe wall biofilms due to cell death or flow dynamics could release the entrapped Fe and Mn into the bulk water, which could lead to a discoloured water event. Hence, managing biofilm growth on drinking water pipelines should be considered by water utilities to minimize accumulation of Fe and Mn in distribution networks.
Biocathodic denitrification using bioelectrochemical systems (BES) have shown promise for both wastewater and groundwater treatment. Typically, these systems involve anodic carbon oxidation and cathodic denitrification catalyzed by two electroactive biofilms located separately at an anode and a cathode. However, process efficiencies are often limited by pH drifts in the respective electrode-biofilms: acidification (pH <5.5) in the bioanode and basification (pH >8.5) in the biocathode. Here, we describe for the first time a single electroactive biofilm that acts as a bioanode and a biocathode, alternately catalyzing anodic acetate oxidation (Coulombic efficiency (CE) 85.3%) and cathodic denitrification (CE 87.3%) (-400 mV Ag/AgCl). Our results indicate that the ano-cathodophilic biofilm denitrified autotrophically using the electrode (-200 to -600 mV Ag/AgCl) as a direct electron donor. Further, the alkalinity produced from cathodic denitrification partially (19%) neutralized the acidity of the anodic reaction. Switching the electrode potential to temporarily favor either an anodic or cathodic reaction may represent a unique method for removing carbon and nitrate from contaminated liquors. This study offers new insights into the development of sustainable BES-based nutrient removal processes.
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