Cooperation and competition between denitrification and chromate reduction in a hydrogen-based membrane biofilm reactor.

Competition and cooperation between denitrification and Cr(VI) reduction in a H2-based membrane biofilm reactor (H2-MBfR) were documented over 55 days of continuous operation. When nitrate (5 mg N/L) and chromate (0.5 mg Cr/L) were fed together, the H2-MBfR maintained approximately 100 % nitrate removal and 60 % chromate Cr(VI) removal, which means that nitrate outcompeted Cr(VI) for electrons from H2 oxidation. Removing nitrate from the influent led to an immediate increase in Cr(VI) removal (to 92 %), but Cr(VI) removal gradually deteriorated, with the removal ratio dropping to 14 % after five days. Cr(VI) removal resumed once nitrate was again added to the influent. 16S rDNA analyses showed that bacteria able to carry out H2-based denitrification and Cr(VI) reduction were in similar abundances throughout the experiment, but gene expression for Cr(VI)-reduction and export shifted. Functional genes encoding for energy-consuming chromate export (encoded by ChrA) as a means of bacterial resistance to toxicity were more abundant than genes encoding for the energy producing Cr(VI) respiration via the chromate reductase ChrR-NdFr. Thus, Cr(VI) transport and resistance to Cr(VI) toxicity depended on H2-based denitrification to supply energy. With Cr(VI) being exported from the cells, Cr(VI) reduction to Cr(III) was sustained. Thus, cooperation among H2-based denitrification, Cr(VI) export, and Cr(VI) reduction led to sustained Cr(VI) removal in the presence of nitrate, even though Cr(VI) reduction was at a competitive disadvantage for utilizing electrons from H2 oxidation.

Double-edged sword effects of dissimilatory nitrate reduction to ammonium (DNRA) bacteria on anammox bacteria performance in an MBR reactor.

Dissimilatory nitrate reduction to ammonium (DNRA) bacteria imposing double-edged sword effects on anammox bacteria were investigated in an anammox-membrane bioreactor (MBR) experiencing an induced crash-recovery event. During the experiment, the anammox-MBR was loaded with NH4+-N:NO2--N ratios (RatioNH4+-N: NO2--N) of 1.20-1.60. Initially, the anammox-MBR removed over 95% of 100 mg/L NH4+-N and 132 mg/L NO2--N (RatioNH4+-N: NO2--N = 0.76, the well accepted stoichiometric RatioNH4+-N: NO2--N for anammox) in the influent (Stage 0). Then, we induced a system crash-recovery event via nitrite shock loadings to better understand responses from different guilds of bacteria in anammox-MBR, loaded with 1.60 RatioNH4+-N: NO2--N with 100 mg/L NO2--N in the influent (Stage 1). Interestingly, the nitrogen removal by anammox bacteria was maintained for about 20 days before starting to decrease significantly. In Stage 2, we further increased influent nitrite concentration to 120 mg/L (1.33 RatioNH4+-N: NO2--N) to simulate a high nitrite toxicity scenario for a short period of time. As expected, nitrogen removal efficiency dropped to only 16.8%. After the induced system crash, anammox-MBR performance recovered steadily to 93.2% nitrogen removal with a 1.25 RatioNH4+-N:NO2--N and a low nitrite influent concentration of 80 mg/L NO2--N. Metagenomics analysis revealed that a probable causality of the decreasing nitrogen removal efficiency in Stage 1 was the overgrowth of DNRA-capable bacteria. The results showed that the members within the Ignavibacteriales order (21.7%) out competed anammox bacteria (17.0%) in the anammox-MBR with elevated nitrite concentrations in the effluent. High NO2--N loading (120 mg N/L) further caused the predominant Candidatus Kuenenia spp. were replaced by Candidatus Brocadia spp. Therefore, it was evident that DNRA bacteria posed negative effects on anammox with 1.60 RatioNH4+-N: NO2--N. Also, when 120 mg/L NO2--N fed to anammox-MBR (RatioNH4+-N: NO2--N = 1.33), canonical denitrification became the primary nitrogen sink with both DNRA and anammox activities decreased. They probably fed on lysed microbial cells of anammox and DNRA. In Stage 3, a low RatioNH4+-N: NO2--N (1.25) with 80 mg/L NO2--N was used to rescue the system, which effectively promoted DNRA-capable bacteria growth. Although anammox bacteria's abundance was only 7.7% during this stage, they could be responsible for about 90% of the total nitrogen removal during this stage.

Identification of dissimilatory nitrate reduction to ammonium (DNRA) and denitrification in the dynamic cake layer of a full-scale anoixc dynamic membrane bioreactor for treating hotel laundry wastewater.

Identification of dissimilatory nitrate reduction to ammonium (DNRA) and denitrification in the dynamic cake layer of a full-scale anoixc dynamic membrane bioreactor (AnDMBR) for treating hotel laundry wastewater was studied. A series of experiments were conducted to understand the contributions of DNRA and canonical denitrification activities in the dynamic cake layer of the AnDMBR. The dynamic cake layer developed included two phases - a steady transmembrane pressure (TMP) increase at 0.24 kPa/day followed by a sharp TMP jump at 1.26 kPa/day four to five days after the AnDMBR start-up. The nitrogen mass balance results showed that canonical denitrification was predominant during the development of the dynamic cake layer. However, DNRA activity and accumulation of bacteria equipped with a complete DNRA pathway showed a positive correlation to the development of the dynamic cake layer. Our metagenomic analysis identified an approximately 18% of the dynamic cake layer bacterial community has a complete DNRA pathway. Pannonibacter (1%), Thauera (0.8%) and Pseudomonas (3%) contained all genes encoding for funcional enzymes of both DNRA (nitrate reductase and DNRA nitrite reductase) and denitrification (nitrate reductase, nitrous oxide reductase and nitric oxide reductase). No other metagenome-assembled genomes (MAGs) possessed a complete cononical denitrification pathway, indicating food-chain-like interactions of denitrifiers in the dynamic cake layer. We found that COD loading rate could be used to control DNRA and canonical denitrification activities during the dynamic cake layer formation.

Core carbon fixation pathways associated with cake layer development in an anoxic-oxic biofilm-membrane bioreactor treating textile wastewater.

Microbial carbon fixation pathways have not yet been adequately understood for their role in membrane case layer formation processes. Carbon fixation bacteria can play critical roles in either causing or enhancing cake layer formation in some autotrophic-prone anoxic conditions, such as sulfur-cycling conditions. Understanding the microbes capable of carbon fixation can potentially guide the design of membrane biofouling mitigation strategies in scientific ways. Thus, we used meta-omics methods to query carbon fixation pathways in the cake layers of a full-scale anoxic-oxic biofilm-MBR system treating textile wastewater in this study. Based on the wastewater constituents and other properties, such as anoxic conditions, sulfide-reducing and sulfur-oxidizing bacteria could co-exist in the membrane unit. In addition, low-light radiation conditions could also happen to the membrane unit. However, we could not quantify the light intensity or total energy input accurately because the whole experimental setup was a full-scale system. Potentially complete carbon fixation pathways in the cake layer included the Calvin-Benson-Bassham cycle, Wood-Ljungdahl pathway, and the 3-hydroxypropionate bicycle. We discovered that using aeration could effectively inhibit carbon fixation, which resulted in mitigating membrane cake layer development. However, the aeration resulted in the 3-hydroxypropionate bicycle pathway, presumably used by aerobic sulfur-oxidizing prokaryotes, to become a more abundant carbon fixation pathway in the cake layer under aerobic conditions.

Assimilatory and dissimilatory sulfate reduction in the bacterial diversity of biofoulant from a full-scale biofilm-membrane bioreactor for textile wastewater treatment.

Assimilatory and dissimilatory sulfate reduction (ASR and DSR) are the core bacterial sulfate-reducing pathways involved in wastewater treatment. It has been reported that sulfate-reducing activities could happen within biofoulants of membrane bioreactors during wastewater treatment. Biofoulants are mainly microbial products contributing membrane fouling and subsequent rising energy consumption in driving membrane filtration. Biofoulants from a full-scale biofilm-membrane bioreactor (biofilm-MBR) treating textile wastewater were investigated in this study. During a 10-month operation, sulfate concentrations in the effluent of the biofilm-MBR gradually decreased alongside with the creeping up sulfite concentrations when biofoulants were also building up on membrane modules. Sulfide had no apparent increases in the effluent during this period. Metagenomic analysis revealed diverse microbial communities residing in the biofoulants. Further analysis on their genetic traits revealed abundant ASR's and DSR's functional genes. A plethora of sulfate-reduction bacteria (SRB), including the well-known Desulfovibrio, Desulfainum, Desulfobacca, Desulfobulbus, Desulfococcus, Desulfonema, Desulfosarcina, Desulfobacter, Desulfobacula, Desulfofaba, Desulfotigum, Desulfatibacillum, Desulfatitalea, Desulfobacterium, were detected in the biofoulants. They were believed to play some important carbon and sulfur-cycling roles in our study. Based on metagenomic analysis, we also deduced that ASR was a functionally more important sulfate-reducing route because of the high abundance of assimilatory sulfate reductases detected. Also, the "AMP (adenosine monophosphate)→sulfite" step was a key reaction shared by both ASR and DSR in the biofoulant. This step might be responsible for the sulfite accumulation in the biofilm-MBR effluent. Overall, ASR functional genes in the biofoulants were more abundant. But the bacteria possessing complete DSR pathways caused the sulfide production in the biofilm-MBR.

Core nitrogen cycle of biofoulant in full-scale anoxic & oxic biofilm-membrane bioreactors treating textile wastewater.

Core nitrogen cycle within biofoulant in full-scale anoxic & oxic biofilm-membrane bioreactor (bMBR) treating textile wastewater was investigated. Wastewater filtered through membrane with biofoulant had elevated NH4+-N and NO2--N concentrations corresponding to decreased NO3--N concentrations. Nevertheless, total nitrogen concentrations did not change significantly, indicating negligible nitrogen removal activities within biofoulant. Metagenomic analysis revealed a lack of genes, such as AmoCAB and Hao in biofoulant, indicating absence of nitrification or anammox populations. However, genes encoding complete pathway for dissimilatory nitrate reduction to ammonium (DNRA) were discovered in 15 species that also carry genes encoding both nitrate reductase and nitrite reductase. No specie contained all genes for complete denitrification pathway. High temperature, high C:N ratio, and anoxic conditions of textile wastewater could favorite microbes growth with DNRA pathway over those with canonical denitrification pathway. High dissolved oxygen concentrations could effectively inhibit DNRA to minimize ammonia concentration in the effluent.