Prospect of denitrifying anaerobic methane oxidation (DAMO) application on wastewater treatment and biogas recycling utilization.

Old-fashioned wastewater treatments for nitrogen depend on heterotrophic denitrification process. It would utilize extra organic carbon source as electron donors when the C/N of domestic wastewater was too low to ensure heterotrophic denitrification process. It would lead to non-compliance with carbon reduction targets and impose an economic burden on wastewater treatment. Denitrifying anaerobic methane oxidation (DAMO), which could utilize methane serving as electron donors to replace traditional organic carbon (methanol or sodium acetate), supplies a novel approach for wastewater treatment. As the primary component of biogas, methane is an inexpensive carbon source. With anaerobic digestion becoming increasingly popular for sludge reduction in wastewater treatment plants (WWTPs), efficient biogas utilization through DAMO can offer an environmentally friendly option for in-situ biogas recycling. Here, we reviewed the metabolic principle and relevant research for DAMO and biogas recycling utilization, outlining the prospect of employing DAMO for wastewater treatment and biogas recycling utilization in WWTPs. The application of DAMO provides a new focal point for enhancing efficiency and sustainability in WWTPs.

Enhanced methane production in anaerobic digestion: A critical review on regulation based on electron transfer.

Anaerobic digestion (AD) is a potential bioprocess for waste biomass utilization and energy conservation. Various iron/carbon-based CMs (e.g., magnetite, biochar, granular activated carbon (GAC), graphite and zero valent iron (ZVI)) have been supplemented in anaerobic digestors to improve AD performance. Generally, the supplementation of CMs has shown to improve methane production, shorten lag phase and alleviate environmental stress because they could serve as electron conduits and promote direct interspecies electron transfer (DIET). However, the CMs dosage varied greatly in previous studies and CMs wash out remains a challenge for its application in full-scale plants. Future work is recommended to standardize the CMs dosage and recover/reuse the CMs. Moreover, additional evidence is required to verify the electrotrophs involved in DIET.

A novel sulfide-driven denitrification methane oxidation (SDMO) system: Operational performance and metabolic mechanisms.

Microbial denitrification is a crucial biological process for the treatment of nitrogen-polluted water. Traditional denitrification process consumes external organic carbon leading to an increase in treatment costs. We developed a novel sulfide-driven denitrification methane oxidation (SDMO) system that integrates autotrophic denitrification (AD) and denitrification anaerobic methane oxidation (DAMO) for cost-effective denitrification and biogas utilization in situ. Two SDMO systems were operated for 735 days, with nitrate and nitrite serving as electron acceptors, to explore the performance of sewage denitrification and characterize metabolic mechanisms. Results showed SDMO system could reach as high as 100% efficiency of nitrogen removal and biogas desulfurization without an external carbon source when HRT was 10 days and inflow nitrogen concentrations were 50-100 mgN·L-1. Besides, nitrate was a preferable electron acceptor for SDMO system. Biogas not only enhanced nitrogen removal but also intensified the DAMO, nitrogen removed through DAMO contribution doubled as original period from 2.9 mgN·(L·d)-1 to 6.2 mgN·(L·d)-1, and the ratio of nitrate removal through AD to DAMO was 1.2:1 with nitrate as electron acceptor. While nitrogen removed almost all through AD contribution and DAMO was weaken as before, the ratio of nitrate removal through AD to DAMO was 21.2:1 with nitrite as electron acceptor. Biogas introduced into SDMO system with nitrate inspired the growth of DAMO bacteria Candidatus Methylomirabilis from 0.3% to 19.6% and motivated its potentiality to remove nitrate without ANME archaea participation accompanying with gene mfnE upregulating ∼100 times. According to the reconstructed genome from binning analysis, the dramatically upregulated gene mfnE was derived from Candidatus Methylomirabilis, which may represent a novel metabolism pathway for DAMO bacteria to replace the role of archaea for nitrate reduction.

Enhancement of methane production from waste activated sludge using hybrid microbial electrolysis cells-anaerobic digestion (MEC-AD) process - A review.

Hybrid microbial electrolysis cells-anaerobic digestion (MEC-AD) was proved to increase methane productivity and methane yield of waste activated sludge (WAS) by establishing direct interspecies electron transfer method and enriching functional microorganisms. This review first summarized the pretreatment methods of WAS for MEC-AD and then reviewed the reactor configurations, operation parameters, and the economic benefit of MEC-AD. Furthermore, the enhancement mechanisms of MEC-AD were reviewed based on the analysis of thermodynamics and microbial community. It was found that the decrease of hydrogen partial pressure due to the hydrogenotrophic methanogens enriched in cathodic biofilm and direct interspecies electron transfer between exoelectrogens and anode were the core mechanisms for improving acidogenesis, acetogenesis, and methanogenesis. Finally, the potentially technological issues that need to be addressed to increase energy efficiency in large-scale MEC-AD processes were discussed.

Simultaneous removal of NOX and SO2 from flue gas in an integrated FGD-CABR system by sulfur cycling-mediated Fe(II)EDTA regeneration.

Chemical absorption-biological reduction (CABR) process is an attractive method for NOX removal and Fe(II)EDTA regeneration is important to sustain high NOX removal. In this study a sustainable and eco-friendly sulfur cycling-mediated Fe(II)EDTA regeneration method was incorporated in the integrated biological flue gas desulfurization (FGD)-CABR system. Here, we investigated the NOX and SO2 removal efficiency of the system under three different flue gas flows (100 mL/min, 500 mL/min, and 1000 mL/min) and evaluated the feasibility of chemical Fe(III)EDTA reduction by sulfide in series of batch tests. Our results showed that complete SO2 removal was achieved at all the tested scenarios with sulfide, thiosulfate and S0 accumulation in the solution. Meanwhile, the total removal efficiency of NOX achieved ∼100% in the system, of which 3.2%-23.3% was removed in spray scrubber and 76.7%-96.5% in EGSB reactor along with no N2O emission. The optimal pH and S2-/Fe(III)EDTA for Fe(II)EDTA regeneration and S0 recovery was 8.0 and 1:2. The microbial community analysis results showed that the cooperation of heterotrophic denitrifier (Saprospiraceae_uncultured and Dechloromonas) and iron-reducing bacteria (Klebsiella and Petrimonas) in EGSB reactor and sulfide-oxidizing, nitrate-reducing bacteria (Azoarcus and Pseudarcobacter) in spray scrubber contributed to the efficient removal of NOX in flue gas.

The interaction between Pseudomonas C27 and Thiobacillus denitrificans in the integrated autotrophic and heterotrophic denitrification process.

Compared to autotrophic and heterotrophic denitrification process, the integrated autotrophic and heterotrophic denitrification (IAHD) shows wider foreground of applications in the actual wastewaters with organic carbon, nitrogen and sulfur co-existing. The efficient co-removal of sulfur, nitrogen, and carbon in the IAHD system is guaranteed by the interaction between heterotrophic and autotrophic denitrificans. In order to further explore the interaction between functional bacteria, Pseudomonas C27 and Thiobacillus denitrifcans were selected as typical heterotrophic and autotrophic bacteria, and their characteristics metabolic responses to different sulfide concentrations were studied. Pseudomonas C27 had higher metabolic activity than T. denitrificans in the IAHD medium with sulfide concentration of 3.12-15.62 mmol/L. Moreover, the fastest sulfide removal rate (0.35 mmol/L·h) was achieved with a single inoculation of Pseudomonas C27. Meanwhile, in mixed inoculant conditions, the interaction between Pseudomonas C27 and T. denitrificans (P:T = 3:1, P:T = 1:1 and P:T = 1:3) yielded the highest sulfide removal efficiency (more than 85%) when sulfide concentration was 6.25-12.5 mmol/L. Additionally, the sulfide removal rate increased with the inoculation proportion of Pseudomonas C27. Thus, this apparent interaction provided a theoretical basis for further understanding and guidance on the efficient operation of IAHD system.

The underlying mechanism of enhanced methane production using microbial electrolysis cell assisted anaerobic digestion (MEC-AD) of proteins.

Anaerobic digestion (AD) is a promising technology capable of converting waste matter into bio-energy. Recent studies have reported that microbial electrolysis cell assisted anaerobic digestion (MEC-AD) is an effective system for methane production from organic waste, via enhanced electron transfer. However, little is known about the effects of applied voltage on the AD of proteins. Herein, the mechanism of MEC-AD on protein digestion was investigated using varying concentrations of bovine serum albumin (BSA) as the protein substrate (500 mg/L, 4 g/L, and 20 g/L BSA). Experimental results showed that the applied voltage can not only enhance the methane production rate from 23.8% to 45.6% at low and medium organic loading (BSA concentration of 500 mg/L and 4 g/L), but also improve the methanogenesis efficiency increased by 225.4% at high BSA concentration (20 g/L) with the applied voltage of 0.6 V compared to that with open circuit. Mechanism explorations revealed that the applied voltage significantly enhanced the acidogenesis and methanogenesis processes in the AD of proteins. Microbial community characterization showed that with the applied voltage, the abundance of fermentative bacteria increased by 46.7 % at the anode, while, the abundance of Methanobacterium at the cathode increased from 10.4 to 84.3%, indicating the methanogenesis pathway transformed from acetoclastic to hydrogenotrophic. External circuit electron transfer calculations demonstrated that only 10% of the produced methane could be attributed to direct interspecies electron transfer (DIET). From a thermodynamic perspective, the applied external voltage led to a reduction in the cathodic potential to -0.9 V, which is beneficial for enhanced methane production via mediated interspecies electron transfer (MIET) by enrichment of hydrogenotrophic methanogens. The findings reported here reveal the previously unrecognized contribution of proteins to MEC-AD, while also furthering our understanding of the role of applied voltage in the MEC-AD process.

Simultaneous removal of carbon dioxide, sulfur dioxide and nitric oxide in a biofilter system: Optimization operating conditions, removal efficiency and bacterial community.

Anthropogenic NOx, SO2 and CO2 emission from the fossil-fuel-fired power plants has aroused growing attention. This study investigated the removal performance of CO2, SO2 and NOx in flue gas as well as conversion efficiency of nitric- and sulfur-compounds in liquid phase in a biofilter. In order to develop the potential of the biofilter, simulative industry wastewater was employed as the spray solution. The satisfactory flue gas removal performance (75.23% CO2, 100% SO2 and 82.81% NO) were achieved under the optimal operating conditions of biofilter: initial solution pH of 9 and liquid-gas ratio (L/G) of 3. The gas film mass transfer coefficients (kGa) results showed that the resistance of gas mass transfer was decreased with increasing the pH value and L/G ratio, respectively. The final transformation product of NO was mostly N2 while about 78% SO2 was converted to elemental sulfur. The microbial community analysis results showed that the relative abundance of bacteria with denitrification capacity was increased by 3.05% which might have contributed to the conversion of NO intermediates products in present study. Collectively, this biofilter system achieve a better flue gas removal performance via the proper operation system, which provides an economic feasible strategy of flue gas purification and increases potential for industrial application.

Revealing the role of nitrate on sulfide removal coupled with bioenergy production in Chlamydomonas sp. Tai-03: Metabolic pathways and mechanisms.

Recently, simultaneous sulfide removal and bioenergy production by microalgal treatment have attracted growing attention. However, the response of nitrogen metabolism to the sulfide-removal process has yet to be explored. Here, variable levels of sulfide could be completely removed by Chlamydomonas sp. Tai-03 under both high and low nitrate conditions in synthetic wastewaters. The highest sulfide removal rate of 5.56 mg-S L-1 h-1 was achieved with the addition of 100 mg L-1 sulfide in the presence of high nitrate. Meanwhile, sulfide was chemically oxidized to sulfate and then ingested by microalgae. Interestingly, sulfide-removal efficiency critically depended on nitrate concentration. Sulfide can also enhance the ability of microalgae to assimilate nitrogen. Based on the analysis of sulfur- and nitrogen-related metabolic profiling, serine as a precursor decreased by 94 % under low levels of nitrate, which induced the significant inhibition of cysteine and methionine biosynthesis. The results indicated that nitrogen source played a critical role in the sulfur cycle because of the positive relationship between the aforementioned metabolic processes and nitrate concentration. Additionally, sulfide can improve lipid and carbohydrate productivity under high levels of nitrate. This study enhances our understanding of the mechanisms underlying the simultaneous removal of sulfide and alternative bioenergy production.

Heterotrophic sulfide-oxidizing nitrate-reducing bacteria enables the high performance of integrated autotrophic-heterotrophic denitrification (IAHD) process under high sulfide loading.

Micro-aerobic enhancement technology has been developed as an effective tool to enhance simultaneous removal of sulfide, nitrate and organic carbon during the integrated autotrophic-heterotrophic denitrification (IAHD) process under high loading; however, its mechanism of enhancement for functional bacteria remains ambiguous. In this study, we discovered that heterotrophic sulfide-oxidizing nitrate-reducing bacteria (h-soNRB) are responsible for enhancing IAHD performance under micro-aerobic conditions with high sulfide loading. In a continuous IAHD bioreactor, aeration rate of 2.6 mL min-1·L-1 promoted 2 to 4 times higher removal efficiencies of sulfide, nitrate and acetate with an influent sulfide concentration of 18.75 mmol/L. Metagenomic analysis revealed that trace oxygen stimulated the abundance of genes responsible for sulfide oxidation (sqr, glpE, pdo, sox and cysK), which were upregulated by 15.2%-129.9%, and the genes encoding nitrate reductase were up-regulated by 67.4%. The increased acetate removal efficiency was attributed to upregulation of ack, pta and TCA cycle related genes. The h-NRB Pseudomonas, Azoarcus, Thauera and Halomonas were detected and regarded as h-soNRB in our bioreactor. According to Illumina MiSeq sequencing, these genera were absolutely dominant in the micro-aerobic microbial community at relative abundances ranging from 82.72% to 90.84%. The sulfide, nitrate and acetate removal rates of Pseudomonas C27, a typical h-soNRB, were at least 10 times higher under micro-aerobic conditions than under anaerobic conditions. Besides, the sulfur, nitrogen and carbon metabolic network was constructed based on the Pseudomonas C27 genome. The pdo and cysK genes found in this strain may be the most advantageous for autotrophic sulfide oxidizing nitrate reducing bacteria (a-soNRB), which are closely related to the high-efficiency sulfide, nitrate and acetate removal performance under high sulfide concentrations and a limited oxygen supply. In addition, after micro-aerobic cultivation, the anaerobic sulfide loading tolerance of the IAHD bioreactor increased from 18.75 to 37.5 mmol/L with sulfide, nitrate and acetate removal efficiencies increasing 1.5 to 3 times, which suggests that intermittent micro-aeration might be a more economical and efficient regime for high-sulfide IAHD regulation.

The stimulating metabolic mechanisms response to sulfide and oxygen in typical heterotrophic sulfide-oxidizing nitrate-reducing bacteria Pseudomonas C27.

Micro-aeration is an effective tool that helps integrated autotrophic and heterotrophic denitrification process to withstand high sulfide concentration by making heterotrophic sulfide-oxidizing nitrate-reducing bacteria (h-soNRB) prevail. For further understanding of the dominance of h-soNRB, Pseudomonas C27 was selected as the typical bacterium and its metabolic characteristics responding to sulfide and oxygen stimulation were studied. Under high sulfide concentration condition, addition of trace oxygen led to a two-stage sulfide oxidation process, and sulfide oxidation rate in the first stage was 1.4 times more than that under anaerobic condition. According to transcriptome analysis, the pdo gene significantly up-regulated 2.36 and 2.57 times with and without oxygen under stimulation of high sulfide concentration. Additionally, two possible enhanced sulfide removal pathways coping with high sulfide concentration, namely sqr-cysI-gpx-gor-glpE and cysK-gshA-gshB-pdo-glpE, caused by oxygen were proposed in Pseudomonas C27. These findings provide a theoretical basis for locating high-efficiency sulfur oxidase in h-soNRB.

Bioelectrochemical system for the enhancement of methane production by anaerobic digestion of alkaline pretreated sludge.

An increasing interest is devoted to combined microbial electrolysis cell-anaerobic digestion (MEC-AD) system which could convert waste activated sludge into biogas. In this study series tests were initially conducted to study the effect of alkaline pretreatment on AD system and the results showed that alkaline pretreatment could promote the dissolution of organic matters in the sludge and thus improve the methane production. Then, the methane production in combined MEC-AD system fed with alkaline-pretreated sludge was investigated. The results indicated that the methane productions increased by 37% and 42% when applied voltage was 0.5 V and 0.8 V. The microbial electrochemical system strongly promoted the growth of Euryarchaeota (Methanosaeta and Methanobacterium). Meanwhile, the abundance of Paraclostridium increased from 17.9% to 38.5% when applied voltage was 0.8 V, suggesting an enhanced fermentation and acetogenesis process. The results of energy balance estimation indicated that MEC-AD system at 0.5 V could achieve higher net energy output.

Performance of a novel IAHD-DSR process with methane and sulfide as co-electron donors.

A novel integrated autotrophic and heterotrophic denitrification- denitrifying sulfide removal (IAHD-DSR) process was established in this study for biogas desulfurization to simultaneously remove nitrogen in wastewater. The study demonstrated that the system could utilize methane and sulfide as co-electron donors to replace organic carbon source in IAHD process. Three batch tests (B1, B2 and B3) were set up with IAHD sludge to explore how the novel process works. According to mass balance in B2, methane oxidation and sulfide oxidation contributed 18.75 % and 71.25 % to nitrate removal, respectively; however, the contribution of methane oxidation to total nitrogen (TN) removal reached 84.36 %. Sulfide was mainly responsible for the reduction of nitrate to nitrite, while the methane was for nitrite to nitrogen gas in the presence of insufficient sulfide as electron donors. The TN removal in B2 was almost the same as in normal IAHD-DSR process B3-C. The functional genes mcrA and pmoA responsible for methane oxidation were detected in all three batches, with the abundance of 2.23 ×106 copies/(g dry soil) for mcrA in B1 being the highest in three batches. The sulfide addition in B2 increased the abundance of gene pmoA, indicating the enhancement of nitrite reduction coupled with methane oxidation.

[Electron Equilibrium Analysis of Integrated Autotrophic and Heterotrophic Denitrification Process Under Micro-aerobic Conditions].

The integrated autotrophic and heterotrophic denitrification (IAHD) process, which can simultaneously degrade sulfide, nitrate, and organic carbon with nitrate as a solo electron acceptor, has gained increasing attention as a key unit in industrial wastewater treatment. Micro-aerobic technology, which introduces trace oxygen as an additional electron acceptor, has been demonstrated as an effective strategy for enhancing the IAHD performance. This study focus on the electronic balance calculation of the IAHD process and reveals for the first time that the IAHD process can efficient proceed with an insufficient supply of electron acceptors (nitrate) under micro-aerobic conditions. In the IAHD batch tests, the highest sulfide, nitrate, and acetate removal efficiencies and rates were obtained with an electronic deletion rate peak at 55.1%. Further sulfide oxidizing batch tests demonstrated that the electronic deletion rates were 18.7% and 38.2% under oxygen contents of 5 mL and 10 mL, respectively, in the biological sulfide oxidizing process. Illumina sequencing was used to analyze the microbial community structure in the sulfide oxidation process and indicated Thiobacillus, Thauera, Mangroviflexus, and Erysipelothrix dominated in all community compositions, in which the relative abundance of Thiobacillus increased with an increase in the electronic deletion rate. This study reveals a potential linkage between the electronic gap and the enhanced IAHD performance, which proves new insights into the simultaneous sulfur, nitrogen, and organic carbon removal process.

The microbial zonation of SRB and soNRB enhanced the performance of SR-DSR process under the micro-aerobic condition.

The micro-aerobic condition has proven to effectively enhance the COD removal and elemental sulfur (S0) transformation rate in the sulfate reduction-denitrifying sulfide removal (SR-DSR) process. However, the mechanisms of how micro-aerobic condition enhances S0 transformation remain largely unknown. Therefore in this work an integrated investigation was performed to document the mechanisms and the effect of different startup modes (micro-aerobic startup (termed as mSR-DSR) and anaerobic startup (termed as aSR-DSR)) on bioreactor performance and microbial community dynamics. The results showed that micro-aerobic startup achieved a shorter period to reach a stable performance for SR-DSR, which could be one of the factors affecting the choice of the bioreactor startup mode considering engineering application. For all the tested conditions, removal of nitrate, sulfate and lactate were 100%, >80% and 100%, respectively. The maximum transformation rate of elemental sulfur in mSR-DSR was 57%, which was higher than that in aSR-DSR. The mechanism explorations revealed that micro-aerobic condition not only particularly enriched the sulfide-oxidizing, nitrate-reducing bacteria (soNRB) but also promoted the microbial zonation of sulfate-reducing bacteria (SRB) and soNRB, thereby achieving more S0 transformation in the effluent. Under micro-aerobic condition, SRB were mainly distributed in the bottom and middle part of the reactor, while soNRB were assembled in the top. The relative abundance of soNRB in both aSR-DSR and mSR-DSR notably increased to 41.5% and 23.7% at the top when 5 mL air min-1 Lreactor-1 was applied. Furthermore, the degradation of organic carbon was also accelerated under micro-aerobic condition, possibly due to the enrichment of organic compounds degrading bacteria Bacteroidetes_vadin HA17.

Dual purpose microalgae-based biorefinery for treating pharmaceuticals and personal care products (PPCPs) residues and biodiesel production.

Microalgal biotechnologies have emerged with high potential for removal of various organic pollutants, such as pharmaceutical and personal care products (PPCPs), from waste streams. In the present study, the removal mechanisms for three typical PPCPs and the lipid performance of Chlamydomonas sp. Tai-03 were thoroughly investigated. Bisphenol A (BPA) and Tetracycline (TCY) achieved complete removal while only ~20% Sulfamethoxazole (SMX) could be removed, even at low concentrations of 1 mg L-1. The mechanisms of elimination showed variation as only SMX could be removed through biodegradation, while ~68.2% TCY and ~14% BPA were removed by a combination of photolysis and hydrolysis. Analysis revealed three intermediates of SMX biodegradation, two of which exhibited high toxicity. Moreover, the lipid content of Chlamydomonas sp. Tai-03 increased from 5 to 49.5% with the addition of SMX, TCY and BPA, with lipid quality varying according to the type of PPCPs. In particular, the dominant component (C18:1) abundance was increased by 15.2% at 10 mg L-1 TCY. Overall, these findings provide a baseline for optimization of microalgal biodiesel production coupled with efficient PPCPs treatment biotechnology.

Bioreactor performance and microbial community analysis of autotrophic denitrification under micro-aerobic condition.

Autotrophic denitrification process is an effective strategy for treating sulfide and nitrate-enriched wastewater with low organic carbon. This study determined the sulfide oxidation and nitrate reduction rates and characterized the dominant bacteria and microbial community structure stimulated by micro-aerobic conditions in autotrophic denitrification system. With gradually increased sulfide concentration, the sulfide removal rate decreased to 37.8% at S2- = 600 mg/L, while the peak sulfide and nitrate removal rates (100% and 53.8%) were achieved at S2- = 800 mg/L with the air aeration rate of 20 mL/min. The Illumina sequencing results indicated that Thiobacillus accounted for 63% of total operational taxonomic units at generic level with sulfide concentration of 200 mg/L under anaerobic condition. However, Azoarcus, Thauera and Aliidiomorina became the dominant genera under micro-aerobic condition and their abundance significantly and positively related to the sulfide concentration and aeration rate (p < 0.05). According to the 16S metaproteomics functional composition prediction, one potential mechanism for autotrophic denitrifying under micro-aerobic condition was deduced that the oxidation of sulfide to thiosulfate further to sulfite was reinforced by trace oxygen, while the sulfite reductase activity was restrained. The decreased sulfide concentration weakened the toxicity inhibition on denitrifiers and accordingly the performance of autotrophic denitrification process was enhanced by micro-aerobic condition.

Response of the reactor performance and microbial community to a shift of ISDD process from micro-aerobic to anoxic condition.

Micro-aerobic condition has proven to be effective in enhancing sulfide oxidation to elemental sulfur (S0) during integrated simultaneous desulfurization and denitrification process (ISDD). In this study we investigated and compared the performance and microbial community of ISDD process operating under initially anoxic, then micro-aerobic and finally switch back to anoxic condition. For all the three tested scenarios, comparable bioreactor performance in terms of sulfate (95.0 ±â€¯4.4%, 90.6 ±â€¯3.8%, 89.8 ±â€¯3.5%) and nitrate (∼100%) removal was achieved. However, a shift of ISDD bioreactor from micro-aerobic to anoxic environment clearly increased the S0 production (30.6%), relative to that at initial anoxic condition (14.2%). Further anoxic bioreactor operation with different influent nitrate concentrations also obtained satisfactory performance particularly in terms of S0 production. Microbial community analysis results showed that functional microorganisms selectively enriched at micro-aerobic condition, particularly sulfide-oxidizing bacteria (SOB), could also function well and enhance S0 production when bioreactor switching from micro-aerobic to anoxic environment. We proposed that micro-aerobic strategy could function as a bio-selector and provide a new idea in functional microorganisms selectively enrichment for wastewater treatment.

Interactions of functional bacteria and their contributions to the performance in integrated autotrophic and heterotrophic denitrification.

Compared to autotrophic and heterotrophic denitrification process, the Integrated autotrophic and heterotrophic denitrification (IAHD) has wider foreground of applications in the condition where the organic carbon, nitrate and inorganic sulfur compounds usually co-exist in the actual wastewaters. As the most well-known IAHD process, the denitrifying sulfide removal (DSR) could simultaneously convert sulfide, nitrate and organic carbon into sulfur, dinitrogen gas and carbon dioxide, respectively. Thus, systematical metabolic functions and contributions of autotrophic and heterotrophic denitrifiers to the IAHD-DSR performance became an problem demanding to be promptly studied. In this work, three upflow anaerobic sludge bioreactors (UASBs) were individually started up in autotrophic (a-DSR), heterotrophic (h-DSR) and mixotrophic conditions (m-DSR). Then, the operating conditions of each bioreactor were switched to different trophic conditions with low and high sulfide concentrations in the influent (200 and 400 mg/L). The removal efficiencies of sulfide, nitrate and acetate all reached 100% in all three bioreactors throughout the operational stages. However, the sulfur transformation ratio ranged from 34.5% to 39.9% at the low sulfide concentration and from 76.8% to 86.7% at the high sulfide concentration in the mixotrophic conditions. Microbial community structure analyzed by the Illumina sequencing indicated that Thiobacillus, which are autotrophic sulfide-oxidizing, nitrate-reducing bacteria (a-soNRB), was the dominant genus (81.3%) in the a-DSR bioreactor. With respect to the mixotrophic conditions, at low sulfide concentration in the m-DSR bioreactor, Thiobacillus (a-soNRB) and Thauera, which are heterotrophic nitrate-reducing bacteria (hNRB), were the dominant genera, with percentages of 48.8% and 14.9%, respectively. When the sulfide concentration in the influent was doubled, the percentage of Thiobacillus decreased by approximately 9-fold (from 48.8% to 5.4%), and the total percentage of Azoarcus and Pseudomonas, which are heterotrophic sulfide-oxidizing, nitrate-reducing bacteria (h-soNRB), increased by approximately 6-fold (from 10.1% to 59.4%). Therefore, the following interactions between functional groups and their functional mechanisms in the DSR process were proposed: (1) a-soNRB (Thiobacillus) and hNRB (Thauera) worked together to maintain the performance under the low sulfide concentration; (2) h-soNRB (Azoarcus and Pseudomonas) took the place of a-soNRB and worked together with hNRB (Thauera and Allidiomarina) under the high sulfide concentration; and (3) a-soNRB (such as Thiobacillus) were possibly the key bacteria and may have contributed to the low sulfur transformation, and h-soNRB may be responsible for the high sulfur transformation in the DSR process.