Quorum sensing responses of r-/K-strategists Nitrospira in continuous flow and sequencing batch nitrifying biofilm reactors.

A better understanding of r-/K-strategists nitrifiers will help to balance the design and operation of bioprocesses for efficient pollution removal from wastewater. The objectives of study were to investigate the nitrite oxidation biokinetics, biofilm property, microbial community and quorum sensing (QS) of nitrifying biofilm in a continuously flow reactor (CFR) and a sequencing batch reactor (SBR). Results showed that nitrite-oxidizing bacteria were estimated to have a nitrite half saturation constant of 76.23 and 224.73 µM in CFR and SBR, respectively. High-throughput and metagenomic sequencing results showed that Nitrospira and Candidatus Nitrospira defluvii were the dominated nitrite-oxidizing taxa performing nitrite oxidation in both reactors. Nitrifying biofilm developed in CFR and SBR showed obviously different properties. Biofilm in SBR had an obviously higher ratio of polysaccharide and protein in extracellular polymeric substances, and higher thickness than in CFR. Metagenomics and chemical analysis revealed various types of acyl-homoserine lactone (AHL) circuit genes (e.g., luxI, lasI, hdtS) and four types of AHL signaling substances (e.g., C6-HSL, C8-HSL, C10-HSL and 3-oxo-C10-HSL) in nitrifying biofilm. The concentrations of these AHLs in biomass and water phases were obviously higher in SBR than that in CFR. Together, AHLs-based QS might affect the formation of nitrifying biofilm and thus contribute to the different biokinetics of Nitrospira in CFR and SBR. Our insights may reveal the molecular mechanism of Nitrospira for different biokinetics, and indicate the AHL association with Nitrospira adaptation to various conditions.

Effects of ferroferric oxide on propionate methanogenesis in sequencing batch reactors: Microbial community structure and metagenomic analysis.

This study investigated the effects of ferroferric oxide (Fe3O4) on propionate methanogenesis in anaerobic sequencing batch reactor (ASBR). Compared to ASBRC (without Fe3O4 addition), the addition of 10 g/L Fe3O4 (ASBRFe) decreased the maximum methane production rate by 69.6 % when propionate was used as the sole substrate. The addition of Fe3O4 reduced the contents of humic substances, riboflavin and nicotinamide adenine dinucleotide in extracellular polymeric substances. Therefore, Fe3O4 inhibited interspecies electron transfer of microorganisms through electronic mediators. Microbial community analysis revealed that Fe3O4 addition increased the relative abundance of acetate oxidizing bacterium (Mesotoga), but decreased the abundance of hydrogenotrophic methanogen (Methanobacterium). Further metagenomics analysis indicated that Fe3O4 increased the abundance of acetate oxidation genes and decreased that of hydrogenotrophic methanogenesis, quorum sensing and V/A-type ATPase genes. Thus, Fe3O4 reduced propionate methanogenesis during anaerobic digestion. The overall results indicate that Fe3O4 addition inhibits methanogenesis for treatment of propionate-contaminated wastewater in ASBR.

Analyzing the effect of organic carbon on partial nitrification-anammox process based on metagenomics and quorum sensing.

The effects of adding organic carbon on the performance of different partial nitrification-anammox (PNA) process (the activated sludge process and biofilm process) were studied, especially nitrogen removal, functional microbial activity, and microbial community structure. The potential influences of quorum sensing (QS) on the nitrogen metabolism were also analyzed. The results showed that the addition of organic carbon in biofilm systems could reduce total nitrogen (TN) removal percentages, while in activated systems it could increase TN removal percentages. The TN removal percentages in SBBR-CN (the biofilm system with addition of organic carbon) and SBR-CN (the activated sludge system with addition of organic carbon) were 15% and 45%, respectively, and those in SBBR-N (the biofilm system without addition of organic carbon) and SBR-N (the activated sludge system without addition of organic carbon) were 75% and 21%, respectively. Batch experiments have proved that organic carbon inhibited the activities of nitrite-oxidizing bacteria (NOB) and anaerobic ammonia oxidation (anammox) bacteria, and organic carbon could promote the activity of denitrifying bacteria in activated sludge systems. Compared with activated sludge systems, biofilm systems could protect the activity of anammox bacteria. The relative abundances of ammonia oxidizing bacteria (AOB) and anammox bacteria were decreased, while the relative abundances of denitrifying bacteria (Thauera) were increased with the addition of organic carbon. The biofilm systems were more conducive to the growth of anammox bacteria. Metagenomics revealed that the same bacteria might be involved in different nitrogen metabolism, and nitrogen metabolism was achieved through the complex cooperation among functional bacteria. Besides, functional bacteria involving in the nitrogen metabolism had genes related to QS, indicating that QS might affect the nitrogen metabolism by regulating the functional bacteria activity. PRACTITIONER POINTS: PNA was achieved through SBBR and complete nitrification was achieved through SBR under the low ammonia nitrogen concentration condition. The effect of organic carbon on biofilm and activated sludge PNA process was different under the low ammonia nitrogen concentration condition. QS and QQ may affect the nitrogen removal performance by regulating the expression of nitrogen metabolism-related genes.

Coupled effects of ferroferric oxide supplement and ethanol co-metabolism on the methanogenic oxidation of propionate.

Direct interspecies electron transfer (DIET) is a new electron-transfer strategy for enhanced propionate degradation. Ethanol can enrich the DIET species of Geobacter and conductive ferroferric oxide (Fe3O4) can promote DIET. Therefore, coupled effects of ethanol and Fe3O4 on propionate degradation were investigated. The maximum CH4 production rate was increased by 81.4% by adding Fe3O4 when simultaneously fed with ethanol and propionate, while the improvement could not be observed without ethanol. The sludge conductivity and the electron transfer system activity by adding Fe3O4 were increased by 2.66 and 2.73 times, respectively. Besides, the relative abundance of functional microbes such as Geobacter, Syntrophobacter, Smithella, and Methanosaeta, and their functional genes were increased by the supplement of Fe3O4. The improvement of propionate degradation by adding Fe3O4 was largely attributed to the co-existence of ethanol degradation. The DIET between Geobacter and Methanosaeta might provide more energies or rapidly consume the oxidation products to promote the propionate degradation.

Microbial interactions regulated by the dosage of ferroferric oxide in the co-metabolism of organic carbon and sulfate.

Effects of ferroferric oxide (Fe3O4) and organic carbon on co-metabolism of sulfate and organic carbon were investigated. With Fe3O4, the degradation of acetate and sulfate was inhibited when fed with acetate, while the degradation of acetate and propionate produced from ethanol was promoted when fed with ethanol. The dominant sulfate reducing bacteria (SRB) of acetate-fed reactors were Desulfobacteraceae (complete oxidizing SRB, CO-SRB) and Desulfurmonas (incomplete oxidizing SRB, IO-SRB). IO-SRBs of Desulfobulbus and Desulfomicrobium were dominant in ethanol-fed reactors. CO-SRB had higher competitiveness than methanogens to utilize acetate, while IO-SRBs might cooperate with methanogens to produce methane when dosed with ethanol and Fe3O4. The dosage of Fe3O4 changed the dominant methanogen from Methanosarcina to Methanosaeta with acetate as the organic carbon, while increased the relative abundance of Methanosaeta with ethanol as the organic carbon.

Effects of carbon source on methanogenic activities and pathways incorporating metagenomic analysis of microbial community.

In this study, the effects of four types of organic compounds (tryptone, acetate/propionate, glucose and ethanol) on methanogenesis, electron transfer processes and microbial community structure were examined. When tryptone and acetate/propionate were used, the dominant methanogenic pathway was aceticlastic methanogenesis and Methanosarcina was the most abundant methanogen. When glucose or ethanol were provided as the external carbon source, the aceticlastic and hydrogenotrophic pathways were utilised simultaneously, and Methanosarcina and Methanobacterium were enriched. However, the reactor fed with glucose was prone to acidification because volatile fatty acids accumulated in the medium, which inhibited methane synthesis. Geobacter was dominant in the reactor fed with ethanol and 45% of genes encoding pili synthesis were attributable to Geobacter, indicating that direct interspecies electron transfer may be a possible mechanism during syntrophic methanogenesis.

Dynamics of intracellular polymers in enhanced biological phosphorus removal processes under different organic carbon concentrations.

Enhanced biological phosphorus removal (EBPR) may deteriorate or fail during low organic carbon loading periods. Polyphosphate accumulating organisms (PAOs) in EBPR were acclimated under both high and low organic carbon conditions, and then dynamics of polymers in typical cycles, anaerobic conditions with excess organic carbons, and endogenous respiration conditions were examined. After long-term acclimation, it was found that organic loading rates did not affect the yield of PAOs and the applied low organic carbon concentrations were advantageous for the enrichment of PAOs. A low influent organic carbon concentration induced a high production of extracellular carbohydrate. During both anaerobic and aerobic endogenous respirations, when glycogen decreased to around 80 ± 10 mg C per gram of volatile suspended solids, PAOs began to utilize polyphosphate significantly. Regressed by the first-order reaction model, glycogen possessed the highest degradation rate and then was followed by polyphosphate, while biomass decay had the lowest degradation rate.