Converting kitchen waste into value-added fertilizer using thermophilic semi-continuous composting-biofiltration two-stage process with minimized NH3 emission.

Thermophilic semi-continuous composting (TSC) is effective for kitchen waste (KW) treatment, but large amounts of NH3-rich odorous gas are generated. This study proposes a TSC-biofiltration (BF) two-stage process. Compost from the front-end TSC was used as the packing material in the BF to remove NH3 from the exhaust gas. The BF process was effective in removing up to 83.7 % of NH3, and the NH3 content was reduced to < 8 ppm. Seven days of BF improved the quality of the product from TSC by enhancing the germination index to 134.6 %, 36.5 % higher than that in the aerated-only group. Microbial community analysis revealed rapid proliferation and eventual dominance in the BF of members related to compost maturation and the nitrogen cycle from Actinobacteria, Proteobacteria, Chloroflexi, and Bacteroidetes. The results suggest that the TSC-BF two-stage process is effective in reducing NH3 emissions from TSC and improving compost quality.

Anaerobic digestion integrated with microbial electrolysis cell to enhance biogas production and upgrading in situ.

Anaerobic digestion (AD) is an effective and applicable technology for treating organic wastes to recover bioenergy, but it is limited by various drawbacks, such as long start-up time for establishing a stable process, the toxicity of accumulated volatile fatty acids and ammonia nitrogen to methanogens resulting in extremely low biogas productivities, and a large amount of impurities in biogas for upgrading thereafter with high cost. Microbial electrolysis cell (MEC) is a device developed for electrosynthesis from organic wastes by electroactive microorganisms, but MEC alone is not practical for production at large scales. When AD is integrated with MEC, not only can biogas production be enhanced substantially, but also upgrading of the biogas product performed in situ. In this critical review, the state-of-the-art progress in developing AD-MEC systems is commented, and fundamentals underlying methanogenesis and bioelectrochemical reactions, technological innovations with electrode materials and configurations, designs and applications of AD-MEC systems, and strategies for their enhancement, such as driving the MEC device by electricity that is generated by burning the biogas to improve their energy efficiencies, are specifically addressed. Moreover, perspectives and challenges for the scale up of AD-MEC systems are highlighted for in-depth studies in the future to further improve their performance.

Response mechanisms of different Saccharomyces cerevisiae strains to succinic acid.

BACKGROUND: The production of succinic acid (SA) from biomass has attracted worldwide interest. Saccharomyces cerevisiae is preferred for SA production due to its strong tolerance to low pH conditions, ease of genetic manipulation, and extensive application in industrial processes. However, when compared with bacterial producers, the SA titers and productivities achieved by engineered S. cerevisiae strains were relatively low. To develop efficient SA-producing strains, it's necessary to clearly understand how S. cerevisiae cells respond to SA. RESULTS: In this study, we cultivated five S. cerevisiae strains with different genetic backgrounds under different concentrations of SA. Among them, KF7 and NBRC1958 demonstrated high tolerance to SA, whereas NBRC2018 displayed the least tolerance. Therefore, these three strains were chosen to study how S. cerevisiae responds to SA. Under a concentration of 20 g/L SA, only a few differentially expressed genes were observed in three strains. At the higher concentration of 60 g/L SA, the response mechanisms of the three strains diverged notably. For KF7, genes involved in the glyoxylate cycle were significantly downregulated, whereas genes involved in gluconeogenesis, the pentose phosphate pathway, protein folding, and meiosis were significantly upregulated. For NBRC1958, genes related to the biosynthesis of vitamin B6, thiamin, and purine were significantly downregulated, whereas genes related to protein folding, toxin efflux, and cell wall remodeling were significantly upregulated. For NBRC2018, there was a significant upregulation of genes connected to the pentose phosphate pathway, gluconeogenesis, fatty acid utilization, and protein folding, except for the small heat shock protein gene HSP26. Overexpression of HSP26 and HSP42 notably enhanced the cell growth of NBRC1958 both in the presence and absence of SA. CONCLUSIONS: The inherent activities of small heat shock proteins, the levels of acetyl-CoA and the strains' potential capacity to consume SA all seem to affect the responses and tolerances of S. cerevisiae strains to SA. These factors should be taken into consideration when choosing host strains for SA production. This study provides a theoretical basis and identifies potential host strains for the development of robust and efficient SA-producing strains.

High synthetic cost-amino acids reduce member interactions of acetate-degrading methanogenic microbial community.

Introduction: The cooperation among members of microbial communities based on the exchange of public goods such as 20 protein amino acids (AAs) has attracted widespread attention. However, little is known about how AAs availability affects interactions among members of complex microbial communities and the structure and function of a community. Methods: To investigate this question, trace amounts of AAs combinations with different synthetic costs (low-cost, medium-cost, high-cost, and all 20 AAs) were supplemented separately to acetate-degrading thermophilic methanogenic reactors, and the differences in microbial community structure and co-occurring networks of main members were compared to a control reactor without AA supplementation. Results: The structure of the microbial community and the interaction of community members were influenced by AAs supplementation and the AAs with different synthetic costs had different impacts. The number of nodes, links, positive links, and the average degree of nodes in the co-occurrence network of the microbial communities with AAs supplementation was significantly lower than that of the control without AAs supplementation, especially for all 20 AAs supplementation followed by the medium- and high-cost AAs supplementation. The average proportion of positive interactions of microbial members in the systems supplemented with low-cost, medium-cost, high-cost, all AAs, and the control group were 0.42, 0.38, 0.15, 0.4, and 0.45, respectively. In addition, the ecological functions of community members possibly changed with the supplementation of different cost AAs. Discussion: These findings highlight the effects of AAs availability on the interactions among members of complex microbial communities, as well as on community function.

Multi-omics association study of hexadecane degradation in haloarchaeal strain Halogranum rubrum RO2-11.

Haloarchaea with the capacity to degrade alkanes is promising to deal with petroleum pollution in hypersaline environments. However, only a limited number of haloarchaeal species are investigated, and their pathway and mechanism for alkane degradation remain unclear. In this study, Halogranum rubrum RO2-11, a haloarchaeal strain, verified the ability to degrade kerosene and hexadecane in 184 g/L NaCl, with 53% and 52% degradation rates after 9 and 4 days, respectively. Genome sequencing and gene annotation indicated that strain RO2-11 possesses a complete potential alkane-degrading pathway, of which alkane hydroxylases may include CYP450, AlmA, and LadA. Transcriptome and metabolome analyses revealed that the upregulation of related genes in TCA cycle, lysine biosynthesis, and acetylation may help improve hexadecane degradation. Additionally, an alternative degrading pathway of hexadecane based on dual-terminal ß-oxidation may occur in strain RO2-11. It is likely to be the first report of alkane degradation by the genus Halogranum, which may be helpful for applications of oil-pollution bioremediation under high-salt conditions.

Continuous thermophilic composting of distilled grain waste improved organic matter stability and succession of bacterial community.

Continuous thermophilic composting (CTC) is potentially helpful in shortening the composting cycle. However, its universal effectiveness and the microbiological mechanisms involved are unclear. Here, the physicochemical properties and bacterial community dynamics during composting of distilled grain waste in conventional and CTC models were compared. CTC accelerated the organic matter degradation rate (0.2 vs. 0.1 d-1) and shortened the composting cycle (24 vs. 65 d), mainly driven by the synergism of bacterial genera. Microbial analysis revealed that the abundance of Firmicutes was remarkably improved compared to that in conventional composting, and Firmicutes became the primary bacterial phylum (relative abundance >70 %) during the entire CTC process. Moreover, correlation analysis demonstrated that bacterial composition had a remarkable effect on the seed germination index. Therefore, controlling the composting process under continuous thermophilic conditions is beneficial for enhancing composting efficiency and strengthening the cooperation between bacterial genera.

Metabolism of novel potential syntrophic acetate-oxidizing bacteria in thermophilic methanogenic chemostats.

Acetate is a major intermediate in the anaerobic digestion of organic waste to produce CH4. In methanogenic systems, acetate degradation is carried out by either acetoclastic methanogenesis or syntrophic degradation by acetate oxidizers and hydrogenotrophic methanogens. Due to challenges in the isolation of syntrophic acetate-oxidizing bacteria (SAOB), the diversity and metabolism of SAOB and the mechanisms of their interactions with methanogenic partners are not fully characterized. In this study, the in situ activity and metabolic characteristics of potential SAOB and their interactions with methanogens were elucidated through metagenomics and metatranscriptomics. In addition to the reported SAOB classified in the genera Tepidanaerobacter, Desulfotomaculum, and Thermodesulfovibrio, we identified a number of potential SAOB that are affiliated with Clostridia, Thermoanaerobacteraceae, Anaerolineae, and Gemmatimonadetes. The potential SAOB possessing the glycine-mediated acetate oxidation pathway dominates SAOB communities. Moreover, formate appeared to be the main product of the acetate degradation by the most active potential SAOB. We identified the methanogen partner of these potential SAOB in the acetate-fed chemostat as Methanosarcina thermophila. The dominated potential SAOB in each chemostat had similar metabolic characteristics, even though they were in different fatty-acid-fed chemostats. These novel syntrophic lineages are prevalent and may play critical roles in thermophilic methanogenic reactors. This study expands our understanding of the phylogenetic diversity and in situ biological functions of uncultured syntrophic acetate degraders and presents novel insights into how they interact with methanogens.IMPORTANCECombining reactor operation with omics provides insights into novel uncultured syntrophic acetate degraders and how they perform in thermophilic anaerobic digesters. This improves our understanding of syntrophic acetate degradation and contributes to the background knowledge necessary to better control and optimize anaerobic digestion processes.

Different responses of mesophilic and thermophilic anaerobic digestion of waste activated sludge to PVC microplastics.

The effect of microplastics (MPs) retained in waste activated sludge (WAS) on anaerobic digestion (AD) performance has attracted more and more attention. However, their effect on thermophilic AD remains unclear. Here, the influence of polyvinyl chloride (PVC) MPs on methanogenesis and active microbial communities in mesophilic (37 °C) and thermophilic (55 °C) AD was investigated. The results showed that 1, 5, and 10 mg/L PVC MPs significantly promoted the cumulative methane yield in mesophilic AD by 5.62%, 7.36%, and 8.87%, respectively, while PVC MPs reduced that in thermophilic AD by 13.30%, 18.82%, and 19.99%, respectively. Moreover, propionate accumulation was only detected at the end of thermophilic AD with PVC MPs. Microbial community analysis indicated that PVC MPs in mesophilic AD enriched hydrolytic and acidifying bacteria (Candidatus Competibacter, Lentimicrobium, Romboutsia, etc.) together with acetoclastic methanogens (Methanosarcina, Methanosaeta). By contrast, most carbohydrate-hydrolyzing bacteria, propionate-oxidizing bacterium (Pelotomaculum), and Methanosarcina were inhibited by PVC MPs in thermophilic AD. Network analysis further suggested that PVC MPs significantly changed the relationship of key microorganisms in the AD process. A stronger correlation among the above genera occurred in mesophilic AD, which may promote the methanogenic performance. These results suggested that PVC MPs affected mesophilic and thermophilic AD of WAS via changing microbial activities and interaction.

Enrichment of microbial consortia for MEOR in crude oil phase of reservoir-produced liquid and their response to environmental disturbance.

Developing microbial consortiums is necessary for microbial enhanced oil recovery (MEOR) in heavy crude oil production. The aqueous phase of produced fluid has long been considered an ideal source of microorganisms for MEOR. However, it is recently found that rich microorganisms (including hydrocarbon-degrading bacteria) are present in the crude oil phase, which is completely different from the aqueous phase of produced fluid. So, in this study, the microbial consortia from the crude oil phase of produced fluids derived from four wells were enriched, respectively. The microbial community structure during passage was dynamically tracked, and the response of enriched consortia to successive disturbance of environmental factors was investigated. The results showed the crude oil phase had high microbial diversity, and the original microbial community structure from four wells was significantly different. After ten generations of consecutive enrichment, different genera were observed in the four enriched microbial consortia, namely, Geobacillus, Bacillus, Brevibacillus, Chelativorans, Ureibacillus, and Ornithinicoccus. In addition, two enriched consortia (eG1614 and eP30) exhibited robustness to temperature and oxygen perturbations. These results further suggested that the crude oil phase of produced fluids can serve as a potential microbial source for MEOR.

Multi-omics analysis to reveal key pathways involved in low C/N ratio stress response in Pseudomonas sp. LW60 with superior nitrogen removal efficiency.

In practical engineering, nitrogen removal at low temperatures or low C/N ratios is difficult. Although strains can remove nitrogen well at low temperatures, there is no research on the performance and deep mechanism of strains under low C/N ratio stress. In this study, Pseudomonas sp. LW60 with superior nitrogen removal efficiency under low C/N ratio stress was isolated at 4 °C. With a C/N ratio of 2-10, the NH4+-N removal efficiency was 40.02 %-100 % at 4 °C. Furthermore, the resistance mechanism of Pseudomonas sp. LW60 to low C/N ratio stress was deeply investigated by multi-omics. The results of transcriptome, proteome, and metabolome revealed that the resistance of strain LW60 to low C/N ratio stress was attributed to enhanced central carbon metabolism, amino acid metabolism, and ABC transporters, rather than nitrogen removal pathways. This study isolated a strain with low C/N ratio tolerance and deeply explored its tolerance mechanism by multi-omics.

Efficient nitrogen removal by a novel extreme strain, Pseudomonas reactans WL20-3 under dual stresses of low temperature and high alkalinity: Characterization, mechanism, and application.

Although many studies report the resistance of heterotrophic nitrification-aerobic denitrification (HN-AD) strains to single environmental stress, there is no research on its resistance to dual stresses of low temperature and high alkalinity. A novel bacterium Pseudomonas reactants WL20-3 isolated in this study showed removal efficiencies of 100%, 100%, and 97.76% for ammonium, nitrate, and nitrite, respectively, at 4 °C and pH 11.0. Transcriptome analysis revealed that the resistance of strain WL20-3 to dual stresses was attributed not only to the regulation of genes in the nitrogen metabolic pathway, but also to genes in other pathways such as the ribosome, oxidative phosphorylation, amino acid metabolism, and ABC transporters. Additionally, WL20-3 removed 83.98% of ammonium from actual wastewater at 4 °C and pH 11.0. This study isolated a novel strain WL20-3 with superior nitrogen removal under dual stresses and provided a molecular understanding of its tolerance mechanism to low temperature and high alkalinity.

A novel hypothermic strain, Pseudomonas reactans WL20-3 with high nitrate removal from actual sewage, and its synergistic resistance mechanism for efficient nitrate removal at 4 °C.

Nitrate can be well removed by bacteria at 25-30 °C. However, nitrate removal almost ceases at temperatures lower than 5 °C. In this study, a novel hypothermic strain, Pseudomonas reactans WL20-3 exhibited an excellent aerobic nitrate removal ability at 4 °C. It had high capability for the removal of nitrate, total dissolved nitrogen (TDN), and dissolved organic carbon (DOC) at 4 °C, achieving removal efficiencies of 100%, 87.91%, and 97.48%, respectively. The transcriptome analysis revealed all genes involved in the nitrate removal pathway were significantly up-regulated. Additionally, the up-regulation of ABC transporter genes and down-regulation of respiratory chain genes cooperated with the nitrate metabolism pathway to resist low-temperature stress. In actual sewage, inoculated with WL20-3, the nitrate removal efficiency was found to be 70.70%. Overall, these findings demonstrated the impressive capacity of the novel strain WL20-3 to remove nitrate and provided novel insights into the synergistic resistance mechanism of WL20-3 at low temperature.

Biomethane production and microbial strategies corresponding to high organic loading treatment for molasses wastewater in an upflow anaerobic filter reactor.

Molasses wastewater contains high levels of organic compounds, cations, and anions, causing operational problems for anaerobic biological treatment. In this study, an upflow anaerobic filter (UAF) reactor was employed to establish a high organic loading treatment system for molasses wastewater and further investigated the microbial community dynamics in response to this stressful operation. The biogas production increased with an increase in total organic carbon (TOC) loading rate from 1.0 to 14 g/L/day, and then it decreased with further TOC loading rate addition until 16 g/L/day. The UAF reactor achieved a maximum biogas production of 6800 mL/L/day with a TOC removal efficiency of 66.5% at a TOC loading rate of 14 g/L/day. Further microbial analyses revealed that both the bacterial and archaeal communities developed multiple strategies to maintain stable operation of the reactor at high organic loading (e.g., Proteiniphilum and Defluviitoga maintained high abundances throughout the operation; Tissierella temporarily dominated the bacterial community at TOC loading rates of 8.0 to 14 g/L/day; and multi-trophic Methanosarcina shifted as the dominant methanogen at the TOC loading rates of 8.0 to 16 g/L/day). This study presents insights into a high organic loading molasses wastewater treatment system and the microbial flexibility in methane fermentation in response to process disturbances.

Thermodynamic restrictions determine ammonia tolerance of methanogenic pathways in Methanosarcina barkeri.

Ammonia is a ubiquitous potential inhibitor of anaerobic digestion processes, mainly exhibiting inhibition towards methanogenic activity. However, knowledge as to how ammonia affects the methanogens is still limited. In this study, we cultured a multitrophic methanogen, Methanosarcina barkeri DSM 800, with acetate, H2/CO2, and methanol to evaluate the influence of ammonia on different methanogenic pathways. Aceticlastic methanogenesis was more sensitive to increased ammonia concentrations than hydrogenotrophic and methylotrophic methanogenesis. Theoretical maximum NH3 tolerances of M. barkeri fed with acetate, H2/CO2, and methanol were calculated to be 39.1 ± 9.0, 104.3 ± 7.4, and 85.7 ± 1.0 mg/L, respectively. The order of the ΔG range of M. barkeri under three methanogenic pathways reflected the order of ammonia tolerance of M. barkeri. Our results provide insights into the role of the thermodynamic potential of methanogenesis on the tolerance of ammonia stress; and shed light on the mechanism of ammonia inhibition on anaerobic digestion.

Valorizing kitchen waste to produce value-added fertilizer by thermophilic semi-continuous composting followed by static stacking: Performance and bacterial community succession analysis.

To explore an effective decentralized kitchen waste (KW) treatment system, the performance and bacterial community succession of thermophilic semi-continuous composting (TSC) of KW followed by static stacking (SS) was studied. A daily feeding ratio of 10% ensured stable performance of TSC using an integrated automatic reactor; the efficiencies of organic matter degradation and seed germination index (GI) reached 80.88% and 78.51%, respectively. SS for seven days further promoted the quality of the compost by improving the GI to 91.58%. Alpha- and beta-diversity analyses revealed significant differences between the bacterial communities of TSC and SS. Firmicutes, Actinobacteria, Chloroflexi, Gemmatimonadetes, and Myxococcota were dominant during the TSC of KW, whereas the members of Proteobacteria and Bacteroidetes responsible for product maturity rapidly proliferated during the subsequent SS and ultimately dominated the compost with Firmicutes and Actinobacteria. These results provide new perspectives for decentralized KW treatment using TSC for practical applications.

Response of Microbial Communities on Cathode with Different Potentials in a Single-Chamber Reactor.

Microbes use both organic and inorganic compounds as electron donors, with different electronic potentials, to produce energy required for growth in environments. Conventional studies on the effects of different electron donors on microbial community has been extensively studied with a set cathode potential. However, it remains under-researched how a microbial community response to the different redox potentials in different environments. Here, we incubated a lake sediment in a single-chamber reactor equipped with three working electrodes, i.e., with potentials of - 0.29 V, - 0.05 V versus standard hydrogen electrode and open-circuit, respectively. Results reveal that the structure of bacterial communities was highly similar for all closed-circuit electrodes (- 0.29 V, - 0.05 V), while differing significantly from those on open-circuit electrodes. We also show that specific bacteria were preferentially enriched by different electrode potentials, i.e., Pseudomonas and Rhodobacter preferentially grew on - 0.05 V and - 0.29 V cathode potentials, Azospirillum and Bosea preferentially grew on - 0.05 V; while Ferrovibrio, Hydrogenophaga, Delftia, and Sphingobium preferentially grew on - 0.29 V. In addition, microorganisms selectively enriched on open-circuit electrodes possess higher connectivity and closer relationship than microorganisms selectively enriched on closed-circuit electrode.

Evaluation of physicochemical properties, bacterial community, and product fertility during rice straw composting supplemented with different nitrogen-rich wastes.

This study evaluated the compostability of rice straw as the main feedstock (75 % in dry weight), supplemented with three different nitrogen-rich wastes, namely food waste (FW), dairy manure (DM), and sewage sludge (SS). Organic matter (OM) degradation, maturity and fertility of the end-product, and bacterial community structure during the composting processes were compared. All composting processes generated mature end-product within 51 days. Notably, FW addition was more effective to accelerate rice straw OM degradation and significantly improved end-product fertility with a high yield of Chinese cabbage. The succession of the bacterial community was accelerated with FW supplementation. Genera Geobacillus, Chryseolinea, and Blastocatella were significantly enriched during the composting of rice straw with FW supplementation. Finally, temperature, total nitrogen, moisture, pH, and total carbon were the key factors affecting microorganisms. This study provides a promising alternative method to enhance the disposal of larger amounts of rice straw in a shorter time.

The responses of mesophilic and thermophilic anaerobic digestion of municipal sludge to periodic fluctuation disturbance of organic loading rate.

Fluctuation disturbance of organic loading rate (OLR) is common in actual anaerobic digestion (AD), but its effects on AD of municipal sludge gets little attention. This study investigated the responses of reactor performance and active microbial community in mesophilic and thermophilic AD of municipal sludge before, during and after OLR periodic fluctuation disturbance. The performance of both reactors were similar before and after disturbance although some parameter values changed during the disturbance, which indicated their enough buffer capacity to OLR periodic fluctuation. Different microbial community at RNA level was observed in the two reactors. When the OLR disturbance commenced, the microbial community changed greatly in thermophilic AD. Error and attack tolerance of the microbial network was analyzed in order to learn the response mechanisms to OLR disturbance. The results assisted that the thermophilic microbial community was more vulnerable, but the reactor performance of which could be maintained using the functional redundancy strategy under OLR fluctuation disturbance.

Saccharomyces cerevisiae employs complex regulation strategies to tolerate low pH stress during ethanol production.

BACKGROUND: Industrial bioethanol production may involve a low pH environment caused by inorganic acids, improving the tolerance of Saccharomyces cerevisiae to a low pH environment is of industrial importance to increase ethanol yield, control bacterial contamination, and reduce production cost. In our previous study, acid tolerance of a diploid industrial Saccharomyces cerevisiae strain KF-7 was chronically acclimatized by continuous ethanol fermentation under gradually increasing low-pH stress conditions. Two haploid strains B3 and C3 having excellent low pH tolerance were derived through the sporulation of an isolated mutant. Diploid strain BC3 was obtained by mating these two haploids. In this study, B3, C3, BC3, and the original strain KF-7 were subjected to comparison transcriptome analysis to investigate the molecular mechanism of the enhanced phenotype. RESULT: The comparison transcriptome analysis results suggested that the upregulated vitamin B1 and B6 biosynthesis contributed to the low pH tolerance. Amino acid metabolism, DNA repairment, and general stress response might also alleviate low pH stress. CONCLUSION: Saccharomyces cerevisiae seems to employ complex regulation strategies to tolerate low pH during ethanol production. The findings provide guides for the construction of low pH-tolerant industrial strains that can be used in industrial fermentation processes.

Thermophilic semi-continuous composting of kitchen waste: Performance evaluation and microbial community characteristics.

This study evaluated the feasibility, system stability, and microbial community succession of thermophilic semi-continuous composting of kitchen waste (KW). The results revealed that treatment performance was stable at a 10 % feeding ratio, with an organic matter (OM) degradation efficiency of 81.5 % and seed germination index (GI) of 50.0 %. Moreover, the OM degradation efficiency and GI were improved to 83.4 % and 70.0 %, respectively, by maintaining an optimal compost moisture content (50-60 %). However, feeding ratios of ≥ 20 % caused deterioration of the composter system owing to OM overloading. Microbial community analysis revealed that Firmicutes, Actinobacteria, Chloroflexi, Proteobacteria, and Gemmatimonadetes were dominant. Additionally, moisture regulation significantly increased the Proteobacteria abundance by 57.1 % and reduced the Actinobacteria abundance by 57.8 %. Moreover, network analysis indicated that the bacterial community stability and positive interactions between genera were enhanced by moisture regulation. This information provides a useful reference for practical KW composting treatment in the semi-continuous mode.