Salt stimulates sulfide-driven autotrophic denitrification: Microbial network and metagenomics analyses.

Sulfur autotrophic denitrification (SADN) is a promising biological wastewater treatment technology for nitrogen removal, and its performance highly relies on the collective activities of the microbial community. However, the effect of salt (a prevailing characteristic of some nitrogen-containing industrial wastewaters) on the microbial community of SADN is still unclear. In this study, the response of the sulfide-SADN process to different salinities (i.e., 1.5 % salinity, 0.5 % salinity, and without salinity) as well as the involved microbial mechanisms were investigated by molecular ecological network and metagenomics analyses. Results showed that the satisfactory nitrogen removal efficiency (>97 %) was achieved in the sulfide-SADN process (S/N molar ratio of 0.88) with 1.5 % salinity. In salinity scenarios, the genus Thiobacillus significantly proliferated and was detected as the dominant sulfur-oxidizing bacteria in the sulfide-SADN system, occupying a relative abundance of 29.4 %. Network analysis further elucidated that 1.5 % salinity had enabled the microbial community to form a more densely clustered network, which intensified the interactions between microorganisms and effectively improved the nitrogen removal performance of the sulfide-SADN. Metagenomics sequencing revealed that the abundance of functional genes encoding for key enzymes involved in SADN, dissimilatory nitrate reduction to ammonium, and nitrification was up-regulated in the 1.5 % salinity scenario compared to that without salinity, stimulating the occurrence of multiple nitrogen transformation pathways. These multi-paths contributed to a robust SADN process (i.e., nitrogen removal efficiency >97 %, effluent nitrogen <2.5 mg N/L). This study deepens our understanding of the effect of salt on the SADN system at the community and functional level, and favors to advance the application of this sustainable bioprocess in saline wastewater treatment.

Revealing the comprehensive impact of organic compounds on the partial nitrification-anammox system during incineration leachate treatment: metabolic hierarchy and adaptation.

Organics, as widespread pollutants in high-strength ammonia wastewater, typically exert adverse effects on the performance of partial nitrification-anammox (PNA) systems. However, the in-depth knowledge on how microbial consortia respond to these disturbances remains limited. In this study, we unveiled the evolution of complex organic matter flow and its impact on the metabolic hierarchy and adaptation of microbial consortia, employing multi-omics approaches, i.e., 16S amplicon sequencing, metagenomics, and metabolomics. In a two-stage PNA system sequentially treating synthetic wastewater and incineration leachate over 230 days, partial nitrification stayed stable (nitrite accumulation > 97%) while anammox efficiency dropped (nitrogen removal decreased from 86% to 78%). The phenomenon was revealed to be correlated with the evolution of dissolved organic matter (DOM) and xenobiotic organic compounds (XOCs). In the PN stage, ammonia-oxidizing bacteria (AOB) exhibited excellent adaptability through active metabolic regulation after treating leachate. Numerous heterotrophs proliferated to utilize DOM and XOCs, triggering a "boom" state evident in the glycerophospholipid metabolism. However, in the anammox stage, the competition between carbon fixation and central carbon metabolism within autotrophs and heterotrophs became evident. Increased biosynthesis costs inhibited the central metabolism (specific anammox activity decreased by 66%) and the Wood-Ljungdahl pathway of anammox bacteria (AnAOB) in the presence of recalcitrant organics. Additionally, the degradation of organics was limited, exhibiting a "bust" state. This study revealed the metabolic adaption and susceptibility of AOB and AnAOB in response to organics from the leachate, demonstrating the applicability of the two-stage configuration for treating high-strength wastewater containing abundant and diverse organics.

Sulfur autotrophic denitrification as an efficient nitrogen removals method for wastewater treatment towards lower organic requirement: A review.

The sulfur autotrophic denitrification (SADN) process is an organic-free denitrification process that utilizes reduced inorganic sulfur compounds (RISCs) as the electron donor for nitrate reduction. It has been proven to be a cost-effective and environment-friendly approach to achieving carbon neutrality in wastewater treatment plants. However, there is no consensus on whether SADN can become a dominant denitrification process to treat domestic wastewater or industrial wastewater if organic carbon is desired to be saved. Through a comprehensive summary of the SADN process and extensive discussion of state-of-the-art SADN-based technologies, this review provides a systematic overview of the potential of the SADN process as a sustainable alternative for the heterotrophic denitrification (HD) process (organic carbons as electron donor). First, we introduce the mechanism of the SADN process that is different from the HD process, including its transformation pathways based on different RISCs as well as functional bacteria and key enzymes. The SADN process has unique theoretical advantages (e.g., economy and carbon-free, less greenhouse gas emissions, and a great potential for coupling with novel autotrophic processes), even if there are still some potential issues (e.g., S intermediates undesired production, and relatively slow growth rate of sulfur-oxidizing bacteria [SOB]) for wastewater treatment. Then we present the current representative SADN-based technologies, and propose the outlooks for future research in regards to SADN process, including implement of coupling of SADN with other nitrogen removal processes (e.g., HD, and sulfate-dependent anaerobic ammonium oxidation), and formation of SOB-enriched biofilm. This review will provide guidance for the future applications of the SADN process to ensure a robust-performance and chemical-saving denitrification for wastewater treatment.

Novel insights into carbon nanomaterials enhancing anammox for nitrogen removal: Effects and mechanisms.

Carbon nanomaterials (CNMs) possess the properties including large specific surface area, high porosity, and stable chemical structures, presenting significant application advantages in wastewater treatment. Indeed, CNMs are considered to be added to anammox systems to strengthen anammox function, especially to resolve the challenge of anammox technology, i.e., the slow growth rate of anammox bacteria, as well as its high environmental sensitivity. This paper systematically reviews the promotion effects and mechanisms of CNMs on the nitrogen removal performance of anammox system. Among the zero-, one-, and two-dimensional CNMs, two-dimensional CNMs have best promoting effect on the nitrogen removal performance of anammox system due to its excellent conductivity and abundant functional groups. Then, the promotion effects of CNMs on anammox process are summarized from the perspective of anammox activity and bacteria abundance. Furthermore, CNMs not only enhance the anammox process, but also stimulate the coupling of denitrification pathways with anammox, as well as the improvement of system operational stability (alleviating the inhibitions of low temperature and pH fluctuation), thus contributing to the promoted nitrogen removal performance. Essentially, CNMs are capable of facilitating microbial immobilization and electron transfer, which favor to improve the efficiency and stability of anammox process. Finally, this review highlights the gap in knowledge and future work, aiming to provide a deeper understanding of how CNMs can strengthen the anammox system and provide a novel perspective for the engineering of the anammox process.

Oxidative stress and antioxidant mechanisms of obligate anaerobes involved in biological waste treatment processes: A review.

In-depth understanding of the molecular mechanisms and physiological consequences of oxidative stress is still limited for anaerobes. Anaerobic biotechnology has become widely accepted by the wastewater/sludge industry as a better alternative to more conventional but costly aerobic processes. However, the functional anaerobic microorganisms used in anaerobic biotechnology are frequently hampered by reactive oxygen/nitrogen species (ROS/RNS)-mediated oxidative stress caused by exposure to stressful factors (e.g., oxygen and heavy metals), which negatively impact treatment performance. Thus, identifying stressful factors and understanding antioxidative defense mechanisms of functional obligate anaerobes are crucial for the optimization of anaerobic bioprocesses. Herein, we present a comprehensive overview of oxidative stress and antioxidant mechanisms of obligate anaerobes involved in anaerobic bioprocesses; as examples, we focus on anaerobic ammonium oxidation bacteria and methanogenic archaea. We summarize the primary stress factors in anaerobic bioprocesses and the cellular antioxidant defense systems of functional anaerobes, a consortia of enzymatic and nonenzymatic mechanisms. The dual role of ROS/RNS in cellular processes is elaborated; at low concentrations, they have vital cell signaling functions, but at high concentrations, they cause oxidative damage. Finally, we highlight gaps in knowledge and future work to uncover antioxidant and damage repair mechanisms in obligate anaerobes. This review provides in-depth insights and guidance for future research on oxidative stress of obligate anaerobes to boost the accurate regulation of anaerobic bioprocesses in challenging and changing operating conditions.

Multidisciplinary characterization of nitrogen-removal granular sludge: A review of advances and technologies.

Nitrogen-removal granular sludge (NRGS) is a promising technology in wastewater treatment, with advantages of efficient nitrogen removal, less footprint, lower sludge production and energy consumption, and is a way for wastewater treatment plants to achieve carbon-neutrality. Aerobic granular sludge (AGS) and anammox granular sludge (AnGS) are two typical NRGS technologies that have attracted extensive attention. Mounting evidence has shown strong associations between NRGS properties and the status of NRGS systems; however, a holistic view is still missing. The aim of this article is to provide an overview of NRGS with an emphasis on characterization. Specifically, the integrated nitrogen transformation pathways inside NRGS and the performance of NRGS treating various wastewaters are discussed. NRGS properties are categorized as physical-, chemical-, biological- and systematical ones, presenting current advances and corresponding characterization technologies. Finally, the future prospects for furthering the mechanistic understanding and engineering application of NRGS are proposed. Overall, the technological advancements in characterization have greatly contributed to understanding NRGS properties, which are potential factors for optimizing the performance and evaluating the working status of NRGS. This review will provide guidance in characterizing NRGS properties and boost the introduction of novel characterization technologies.