Cold-resistant performance and the promoted development of functional community with flexible metabolic patterns in a Biofilm Bio-Nutrient Removal (BBNR) system amended with supplementary carbon source for phosphorus recovery.

The need for recovery of phosphorus (P) from wastewater has accelerated the retrofitting of existing bio-nutrient removal (BNR) processes into bio-nutrient removal-phosphorus recovery processes (BNR-PR). A periodical carbon source supplement is needed to facilitate the P-recovery. But the impact of this amendment on the cold resistances of the reactor and the functional microorganisms (for nitrogen and phosphorus (P) removal/recovery) are still unknown. This study presents the performances of a biofilm BNR process with a carbon source regulated the P recovery (BBNR-CPR) process operating at different temperatures. When the temperature was decreased from 25 ± 1 °C to 6 ± 1 °C, the system total nitrogen and total phosphorus removals and the corresponding kinetic coefficients decreased moderately. The indicative genes of the phosphorus-accumulating organisms (e.g., Thauera spp. and Candidatus Accumulibacter spp.) increased significantly. An increase of Nitrosomonas spp. genes aligned to polyhydroxyalkanoates (PHAs), glycine, and extracellular polymeric substance synthesis were observed, which was probably related to cold resistance. The results provide a new vision for understanding the advantages of P recovery-targeted carbon source supplementation for constructing a new type of cold-resistant BBNR-CPR processes.

Dissolved organic matter byproducts from combined low-level alkaline and mechanical treatment of sludge: species, and their bioavailability.

Low-level alkalinity (pH 9-10) coupled with ultrasonic or mechanical cutting with different energy input for obtaining carbon sources were tested for sludge pretreatment process before anaerobic sludge digestion. The differences between the primary sludge (PS) and waste activated sludge (WAS)-derived dissolved organic matter (DOM) species were evaluated for their bioavailability and affinity (in the form of amino acids) to the bio-nutrient removal (BNR) biomass. Soluble microbial by-product-like substances as the predominant DOM components in the raw PS and WAS increased by 23 and 22%, respectively, after low-level alkaline treatment (pH 9-10) and ultrasonication. In addition, the protein components were degraded further as free amino acids (FAAs). The sludge-derived aspartate, glutamate, followed by arginine were the most commonly used FAAs by the BNR biomass. The pattern of recovering this special sludge-derived carbon source to enhance P removal and recovery in the BNR process is depicted.

Optimizing anaerobic digestion: Benefits of mild temperature transition from thermophilic to mesophilic conditions.

Anaerobic digestion (AD) plays a significant role in renewable energy recovery. Upgrading AD from thermophilic (50-57 °C) to mesophilic (30-38 °C) conditions to enhance process stability and reduce energy input remains challenging due to the high sensitivity of thermophilic microbiomes to temperature fluctuations. Here we compare the effects of two decreasing-temperature modes from 55 to 35 °C on cell viability, microbial dynamics, and interspecies interactions. A sharp transition (ST) is a one-step transition by 20 °C d-1, while a mild transition (MT) is a stepwise transition by 1 °C d-1. We find a greater decrease in methane production with ST (88.8%) compared to MT (38.9%) during the transition period. ST mode overproduced reactive oxygen species by 1.6-fold, increased membrane permeability by 2.2-fold, and downregulated microbial energy metabolism by 25.1%, leading to increased apoptosis of anaerobes by 1.9-fold and release of intracellular substances by 2.9-fold, further constraining methanogenesis. The higher (1.6 vs. 1.1 copies per gyrA) metabolic activity of acetate-dependent methanogenesis implied more efficient methane production in a steady mesophilic, MT-mediated system. Metagenomic binning and network analyses indicated that ST induced dysbiosis in keystone species and greatly enhanced microbial functional redundancy, causing loss of microbial syntrophic interactions and redundant metabolic pathways. In contrast, the greater microbial interconnections (average degrees 44.9 vs. 22.1) in MT at a steady mesophilic state suggested that MT could better maintain necessary system functionality and stability through microbial syntrophy or specialized pathways. Adopting MT to transform thermophilic digesters into mesophilic digesters is feasible and could potentially enhance the further optimization and broader application of practical anaerobic engineering.

Functional traits and health implications of the global household drinking-water microbiome retrieved using an integrative genome-centric approach.

The biological safety of drinking water plays a crucial role in public health protection. However, research on the drinking water microbiome remains in its infancy, especially little is known about the potentially pathogenic bacteria in and functional characteristics of the microbiome in household tap water that people are directly exposed to. In this study, we used a genomic-centric approach to construct a genetic catalogue of the drinking water microbiome by analysing 116 metagenomic datasets of household tap water worldwide, spanning nine countries/regions on five continents. We reconstructed 859 high-quality metagenome-assembled genomes (MAGs) spanning 27 bacterial and 2 archaeal phyla, and found that the core MAGs belonging to the phylum Proteobacteria encoded the highest metabolic functional diversity of the 33 key complete metabolic modules. In particular, we found that two core MAGs of Brevibacillus and Methylomona encoded genes for methane metabolism, which may support the growth of heterotrophic organisms observed in the oligotrophic ecosystem. Four MAGs of complete ammonia oxidation (comammox) Nitrospira were identified and functional metabolic analysis suggested these may enable mixotrophic growth and encode genes for reactive oxygen stress defence and arsenite reduction that could aid survival in the environment of oligotrophic drinking water systems. Four MAGs were annotated as potentially pathogenic bacteria (PPB) and thus represented a possible public health concern. They belonged to the genera Acinetobacter (n = 3) and Mycobacterium (n = 1), with a total relative abundance of 1.06 % in all samples. The genomes of PPB A. junii and A. ursingii were discovered to contain antibiotic resistance genes and mobile genetic elements that could contribute to antimicrobial dissemination in drinking water. Further network analysis suggested that symbiotic microbes which support the growth of pathogenic bacteria can be targets for future surveillance and removal.

Microbiome-functionality in anaerobic digesters: A critical review.

Microbially driven anaerobic digestion (AD) processes are of immense interest due to their role in the biovalorization of biowastes into renewable energy resources. The function-versatile microbiome, interspecies syntrophic interactions, and trophic-level metabolic pathways are important microbial components of AD. However, the lack of a comprehensive understanding of the process hampers efforts to improve AD efficiency. This study presents a holistic review of research on the microbial and metabolic "black box" of AD processes. Recent research on microbiology, functional traits, and metabolic pathways in AD, as well as the responses of functional microbiota and metabolic capabilities to optimization strategies are reviewed. The diverse ecophysiological traits and cooperation/competition interactions of the functional guilds and the biomanipulation of microbial ecology to generate valuable products other than methane during AD are outlined. The results show that AD communities prioritize cooperation to improve functional redundancy, and the dominance of specific microbes can be explained by thermodynamics, resource allocation models, and metabolic division of labor during cross-feeding. In addition, the multi-omics approaches used to decipher the ecological principles of AD consortia are summarized in detail. Lastly, future microbial research and engineering applications of AD are proposed. This review presents an in-depth understanding of microbiome-functionality mechanisms of AD and provides critical guidance for the directional and efficient bioconversion of biowastes into methane and other valuable products.

Impacts of organic loading rate and hydraulic retention time on organics degradation, interspecies interactions and functional traits in thermophilic anaerobic co-digestion of food waste and sewage sludge.

This study provided novel insights into the effects of organic loading rate (OLR) and hydraulic retention time (HRT) on thermophilic anaerobic co-digestion of food waste and sewage sludge. The obtained maximum methane (CH4) yield of 328 ± 4 mL CH4/g CODfed at HRT of 15 days (OLR = 5.8 g VS/L/d) was partly attributable to the enhanced acidogenesis, acetogenesis, and methanogenesis phases. The increased key enzyme activities, particularly acetate kinase (improved by 5.2-fold), providing substantial methanogenic substrates for efficient CH4 production. The functional syntrophs that were related to syntrophic decarboxylation, novel acetate oxidation & reductive acetyl-CoA, and ß-oxidation pathways could drive trophic interactions with methanogens. This markedly stimulated hydrogenotrophic Methanoculleus thermophilus metabolism and concomitantly enriched mixotrophic Methanosarcina thermophila. The distinctive cross-feeding interspecies interactions significantly affected the assembly and dynamics of thermophilic consortia. These findings shed light on the physicochemical and microbial mechanisms of HRT- and OLR-dependent enhancement of methanogenesis.

Enhancing methane production in anaerobic co-digestion of sewage sludge and food waste by regulating organic loading rate.

This study presented mechanistic insights into the long-term effects of stepwise-increasing organic loading rates (OLRs) on anaerobic co-digestion (AcoD) of sewage sludge and food waste. The maximum methane (CH4) yield of 500.0 ± 10.5 mL CH4/g VSfed was achieved at medium OLR of 3.5 g VS/L/d. This excellent performance was associated with the high hydrolysis efficiency (78.4%), three-fold enhancement in the acidogenesis enzyme activity, and 87.0% enhanced methanogen activity. Soluble intermediates (carbohydrates and proteins) were largely degraded (>98.5%), especially tyrosine-like and tryptophan-like aromatic proteins. The particulates were effectively decomposed from macromolecules to micromolecules, and the crystallinity of cellulosic substances decreased by 24.5%. The newly-shaped combined syntrophic acetate oxidation-hydrogenotrophic methanogenesis pathway dominated enhanced CH4 production. Energy balance analysis based on medium OLR demonstrated the high energy recovery potential in full-scale AcoD. These findings suggest the optimal medium OLR can facilitate the bioconversion of organics to CH4 through a new metabolic pathway.