Promoting agricultural waste-driven denitrification and nitrogen sequestration with nano-enabled strategy.

Nanotechnology and biotechnology offer promising avenues for bolstering food security through the facilitation of soil nitrogen (N) sequestration and the reduction of nitrate leaching. Nonetheless, a comprehensive and mechanistic evaluation of their effectiveness and safety remains unclear. In this study, a soil remediation strategy employing nano-Fe3O4 and straw in N-contaminated soil was developed to elucidate N retention mechanisms via diverse metagenomics techniques. The findings revealed that subsoil amended with straw, particularly in conjunction with nano-Fe3O4, significantly increased subsoil N content (53.2%) and decreased nitrate concentration (74.6%) in leachate. Furthermore, the enrichment of functional genes associated with N-cycling, sulfate, nitrate, and iron uptake, along with chemotaxis, and responses to environmental stimuli or microbial collaboration, effectively mitigates nitrate leaching while enhancing soil N sequestration. This study introduces a pioneering approach utilizing nanomaterials in soil remediation, thereby offering the potential for the cultivation of safe vegetables in high N input greenhouse agriculture.

Enhanced organic degradation and microbial community cooperation by inoculating Bacillus licheniformis in low temperature composting.

Microbial activity and interaction are the important driving factors in the start-up phase of food waste composting at low temperature. The aim of this study was to explore the effect of inoculating Bacillus licheniformis on the degradation of organic components and the potential microbe-driven mechanism from the aspects of organic matter degradation, enzyme activity, microbial community interaction, and microbial metabolic function. The results showed that after inoculating B. licheniformis, temperature increased to 47.8°C on day 2, and the degradation of readily degraded carbohydrates (RDC) increased by 31.2%, and the bioheat production increased by 16.5%. There was an obvious enhancement of extracellular enzymes activities after inoculation, especially amylase activity, which increased by 7.68 times on day 4. The inoculated B. licheniformis colonized in composting as key genus in the start-up phase. Modular network analysis and Mantel test indicated that inoculation drove the cooperation between microbial network modules who were responsible for various organic components (RDC, lipid, protein, and lignocellulose) degradation in the start-up phase. Metabolic function prediction suggested that carbohydrate metabolisms including starch and sucrose metabolism, glycolysis / gluconeogenesis, pyruvate metabolism, etc., were improved by increasing the abundance of related functional genes after inoculation. In conclusion, inoculating B. licheniformis accelerated organic degradation by driving the cooperation between microbial network modules and enhancing microbial metabolism in the start-up phase of composting.

Screening Ultra-Stable (Phenazine)dioxyalkanocic Acids with Varied Water-Solubilizing Chain Lengths for High-Capacity Aqueous Redox Flow Batteries.

Aqueous redox flow batteries (ARFBs) hold great potential for large-scale energy storage. Recently, research on aqueous flow batteries has shifted toward water-soluble organic molecules with redox capabilities to reduce the use of mineral resources. The chemical and electrochemical stabilities of organic compounds are heavily influenced by their functional groups and reaction sites. In this study, we present a low-cost synthesis of the O-alkyl-carboxylate-functionalized derivatives of 2,3-dihydroxyphenazine, namely, phenazine-(2,3-diyl) dioxy dibutyric acid (DBEP) and phenazine-(2,3-diyl)dioxy diacetic acid (DAEP), which serve as negolytes and exhibit good reversibility and high redox kinetics. The evidence is provided to clarify the capacity degradation mechanisms of DAEP and DBEP by a series of comprehensive characterizations. Similar to anthraquinones functionalized with alkyl chains, the main degradation mechanism of DAEP modified with acetic acid is due to side chain loss. Longer side chains are more stable and can withstand long-term electrochemical reactions. DBEP modified with butyric acid exhibits superior chemical and electrochemical stability. Our results demonstrate that rational molecular design and suitable membranes, such as the alkaline ARFBs based on DBEP negolyte, potassium ferrocyanide (K4Fe(CN)6) posolyte, and custom sulfonated poly(ether ether ketone) membrane, can deliver a high open-circuit voltage of 1.17 V and high capacity retention of 99.997% per cycle for over 1000 cycles at 50 mA cm-2. This study highlights the importance of not only considering the modification position of the molecules but also focusing on the influence of various side chains on the redox core's stability toward sustainable grid-scale energy storage applications.

Synergistic effects of chemical additives and mature compost on reducing H2S emission during kitchen waste composting.

Additives could improve composting performance and reduce gaseous emission, but few studies have explored the synergistic of additives on H2S emission and compost maturity. This research aims to make an investigation about the effects of chemical additives and mature compost on H2S emission and compost maturity of kitchen waste composting. The results showed that additives increased the germination index value and H2S emission reduction over 15 days and the treatment with both chemical additives and mature compost achieved highest germination index value and H2S emission reduction (85%). Except for the treatment with only chemical additives, the total sulfur content increased during the kitchen waste composting. The proportion of effective sulfur was higher with the addition of chemical additives, compared with other groups. The relative abundance of H2S-formation bacterial (Desulfovibrio) was reduced and the relative abundance of bacterial (Pseudomonas and Paracoccus), which could convert sulfur-containing substances and H2S to sulfate was improved with additives. In the composting process with both chemical additives and mature compost, the relative abundance of Desulfovibrio was lowest, while the relative abundance of Pseudomonas and Paracoccus was highest. Taken together, the chemical additives and mature compost achieved H2S emission reduction by regulating the dynamics of microbial community.

Co-inoculation of phosphate-solubilizing bacteria and phosphate accumulating bacteria in phosphorus-enriched composting regulates phosphorus transformation by facilitating polyphosphate formation.

This study aimed to explore the impact of co-inoculating phosphate-solubilizing bacteria (PSB) and phosphate accumulating bacteria (PAB) on phosphorus forms transformation, microbial biomass phosphorus (MBP) and polyphosphate (Poly-P) accumulation, bacterial community composition in composting, using high throughput sequencing, PICRUSt 2, network analysis, structural equation model (SEM) and random forest (RF) analysis. The results demonstrated PSB-PAB co-inoculation (T1) reduced Olsen-P content (1.4 g) but had higher levels of MBP (74.2 mg/kg) and Poly-P (419 A.U.) compared to PSB-only (T0). The mantel test revealed a significantly positive correlation between bacterial diversity and both bioavailable P and MBP. Halocella was identified as a key genus related to Poly-P synthesis by network analysis. SEM and RF analysis showed that pH and bacterial community had the most influence on Poly-P synthesis, and PICRUSt 2 analysis revealed inoculation of PAB increased ppk gene abundance in T1. Thus, PSB-PAB co-inoculation provides a new idea for phosphorus management.

Bacterial wilt suppressive composts: Significance of rhizosphere microbiome.

Composts are often suppressive to several plant diseases, including the devastating bacterial wilt caused by Ralstonia solanacearum. However, the underlying mechanisms are still unclear. Herein, we carried out an experiment with 38 composts collected from different factories in China to study the interlinking among bacterial wilt suppression, the physicochemical properties and bacterial community of the compost, and bacterial community in the rhizosphere of tomato fertilized by compost. Totally 26 composts were suppressive to bacterial wilt, while six composts stimulated the disease. The control efficiency was neither correlated with physicochemical properties (TC, TN, P and K, pH or GI) nor bacterial community of compost, but with rhizosphere bacterial community (r = 0.17, p = 0.016). The control efficiency was also positive correlated with taxa (Rhizobium, Aeromicrobium) known suppressive to R. solanacearum. The mushroom spent or cow manure, from which the two composts were 100% and 77% in control efficiencies against bacterial wilt respectively were subject to a pilot-scale composting reaction. The reproduced composts from mushroom spent or cow manure were only 57% and 23% effective on the control of bacterial wilt, respectively. The analysis of bacterial communities revealed that the relative abundances of R. solanacearum were 28.4% for the control, but only 7.8%-7.9% for compost fertilized tomatoes. The compost from mushroom spent also exerted a strong effect on rhizosphere bacterial community. Taken together, most composts were suppressive to bacterial wilt possibly also by modifying rhizosphere bacterial community towards inhibiting the colonization of R. solanacearum and selecting for beneficial genera of Proteobacteria, Bacteroidetes and Actinobacteria.

Bacillus velezensis BER1 enriched Flavobacterium daejeonense-like bacterium in the rhizosphere of tomato against bacterial wilt.

Beneficial microorganisms can protect crop from phytopathogens, and modify rhizosphere microbiome. However, it is not well-understood whether or how do rhizosphere microorganisms which respond to bioagents contribute to disease suppression. Bacillus velezensis BER1 and tomato bacterial wilt caused by Ralstonia solanacearum were selected as models to disentangle the interactions and mechanisms in the rhizosphere. Bacillus velezensis BER1 greatly suppressed tomato bacterial wilt by over 49.0%, reduced R. solanacearum colonization in the rhizosphere by 36.3%, and significantly enriched two Flavobacterium ASVs (1357 and 2401). A novel colony loop-mediated isothermal amplification (LAMP) assay system was developed to screen out Flavobacterium from tomato rhizosphere bacterial isolates. In vitro tests revealed that cocultivating BER1 with Flavobacterium C45 increased biofilm formation by 18.6%. Climate chamber experiment further revealed that Flavobacterium C45 improved the control efficiency of BER1 on tomato bacterial wilt by 46.0%, decreased the colonization of R. solanacearum in the rhizosphere by 43.1% and elevated the transcription of plant defense gene PR1α in tomato by 45.4%. In summary, Flavobacterium C45 boosted the ability of B. velezensis BER1 to prevent bacterial wilt and the colonization of R. solanacearum, highlighting the importance of helper bacteria on elevating the efficiency of biological control.

Changes in bacterial diversity, co-occurrence pattern, and potential pathogens following digestate fertilization: Extending pathogen management to field for anaerobic digestion of livestock manure.

Digestate can spread pathogens into agroecosystem, posing serious threats to public health. However, the effect of digestate fertilization on digestate- or soil-borne pathogens has not been fully explored. Herein, two settings of microcosm experiment were performed with arable soil and digestate collected at two sites (Beilangzhong or Shunyi) to dissect the succession of the total and potential pathogenic bacterial communities following digestate fertilization. Each experimental setting consisted of three treatments, including digestate aerobically incubated in sterilized soil, and soil amended with sterilized or non-sterilized digestate. Digestate-borne potential pathogenic bacteria were enriched after the aerobic incubation, with Streptococcus sobrinus in the Beilangzhong setting, and Escherichia coli and Enterococcus faecium in the Shunyi setting. Potential soil-borne pathogenic bacteria, such as Acinetobacter lowffii and Pseudomonas fluorescens, were stimulated by the sterilized digestate in the Shunyi setting. Interestingly, S. sobrinus, E. coli, and Ent. faecium did not increase when digestate was amended into the non-sterilized soil, suggesting that soil microorganisms can inhibit the resurgence of these digestate-borne pathogens. A large-scale survey further revealed that organic fertilization exerted a site-dependent effect on different species of potential pathogen, but it did not enrich the total relative abundance of potential pathogenic bacteria in soils. Collectively, these results highlight that pathogen management of anaerobic digestion of livestock manure needs to be extended from anaerobic reactor to field.

Effects of chemical additives and mature compost on reducing nitrogen loss during food waste composting.

This study is aimed at adding different types of mature compost and sulfur powder, as additives into food waste composting to investigate the effect on nitrogen loss and compost maturity. The composting experiment used the in-vessel composting method and was conducted continuously for 15 days. High-throughput sequencing was used to analyze the bacterial community during composting. Results showed that the secondary fermentation mature compost mixed with sulfur powder group had the most reduction of ammonia emission (56%) and the primary fermentation mature compost amendments were the most effective for nitrous oxide emission reduction (37%). The temperature, pH, and nitrogen forms of transformation of the pile significantly affect the nitrogen loss during composting. Firmicutes helped to promote the rapid warming of the pile, and Actinobacteria and Proteobacteria played an important role in decomposition of organic matter. Thermobifida and Ureibacillus had a main contribution to the rapid degradation of organic matter in the process of composting. The relative abundance of nitrogen-fixing bacteria was higher, and the relative abundance of predominantly ammonifying and denitrifying bacteria was lower than the control group, with the addition of different additives.

Role of fungal communities and their interaction with bacterial communities on carbon and nitrogen component transformation in composting with different phosphate additives.

The aim of the study was to compare the succession of fungal community and their interaction with bacterial community during pig manure composting with different phosphate additives and further to identify microbial roles on the transformation of carbon and nitrogen (C&N) components and compost maturity. The results showed that the composition of fungal community was significantly affected by pH in composting and acidic phosphate might postpone the C&N degradation process. Network analysis showed that phosphate additives, especially acidic additives, could increase the interaction of microbial community but acidic phosphate decreased the core fungi:bacteria ratio. Redundancy analysis indicated that the interactions between bacterial and fungal communities played more roles than individual contribution of bacteria or fungi for C&N conversion of composting. Structural equation modeling suggested that bacterial community was positively directly correlated to C&N loss and the participation of fungal community significantly benefited the maturity of composting. pH exhibited a great intermediated role for driving C&N conversion, maturity, and safety of composts by regulating bacterial and fungal community in composting with phosphate addition, which suggested a fast-composting way based on pH regulation by additives.

Insight into the dynamic microbial community and core bacteria in composting from different sources by advanced bioinformatics methods.

Microbial communities are important for high composting efficiency and good quality composts. This study was conducted to compare the changes of physicochemical and bacterial characteristics in composting from different raw materials, including chicken manure (CM), duck manure (DM), sheep manure (SM), food waste (FW), and vegetable waste (VW). The role and interactions of core bacteria and their contribution to maturity in diverse composts were analyzed by advanced bioinformatics methods combined sequencing with co-occurrence network and structural equation modeling (SEM). Results indicated that there were obviously different bacterial composition and diversity in composting from diverse sources. FW had a low pH and different physiochemical characteristics compared to other composts but they all achieved similar maturity products. Redundancy analysis suggested total organic carbon, phosphorus, and temperature governed the composition of microbial species but key factors were different in diverse composts. Network analysis showed completely different interactions of core bacterial community from diverse composts but Thermobifida was the ubiquitous core bacteria in composting bacterial network. Sphaerobacter and Lactobacillus as core genus were presented in the starting mesophilic and thermophilic phases of composting from manure (CM, DM, SM) and municipal solid waste (FW, VW), respectively. SEM indicated core bacteria had the positive, direct, and the biggest (> 80%) effects on composting maturity. Therefore, this study presents theoretical basis to identify and enhance the core bacteria for improving full-scale composting efficiency facing more and more organic wastes.

Risk assessment of three sheep stocking modes via identification of bacterial genomes carrying antibiotic resistance genes and virulence factor genes.

In order to protect the prairie ecological environment, intensive farming has become a prevalent method of sheep stocking. However, the link between captivity stocking mode and ecological risk of sheep feces is still poorly understood. In this study, metagenomics was used to identify the environmental risk of sheep feces among three stocking modes. Our results showed that captivity mode (C) elevated antibiotic resistance in feces, with the abundance of antibiotic resistance genes (ARGs) (5.381 copies/cell) higher than that of half-pen stocking (Fh) (1.093 copies/cell) and grazing mode (Fr) (0.315 copies/cell) (Duncan's test, P < 0.05). Virulence factor genes (VFGs) analysis showed offensive virulence factors had the highest abundance in captivity feces (C: 3.826 copies/cell, Fh: 0.342 copies/cell, Fr: 0.163 copies/cell) (Duncan's test, P < 0.05). 15 metagenome-assembled genomes (MAGs) were identified as potential pathogenic antibiotic resistant bacteria (PARB) and revealed that Escherichia, Klebsiella may be the main host of ARGs and VFGs in sheep feces. Furthermore, the minimal inhibition concentrations (MIC) of tetracycline of E. coli in the captivity feces was 8.6 times and 4.7 times than that of grazing and half-pen stocking samples, respectively. The Non-metric multidimensional scaling (NMDS) revealed that high stocking density leads to feces causing increased harm to the environment. Although feces from sheep raised in captivity and half-pen stocking modes are easier to collect, they are more harmful to the environment and aerobic composting should be done before their application to farmland. This work provides a guideline for better control of the environmental risk of sheep feces from different stocking modes.

Impact of aeration rate on phosphorus conversion and bacterial community dynamics in phosphorus-enriched composting.

This study was to investigate the effects of different aeration rates on phosphorus (P) conversion and bacterial community dynamics in P-enriched composting by 16S rRNA gene sequencing, sequential P fractionation, network analysis and structural equation model (SEM). Results indicated that Olsen P content increased by 138 %, 150 %, 121 % after composting with aeration rate (L kg-1 DM min-1) at 0.2 (AR0.2), 0.4 (AR0.4) and 0.6 (AR0.6). AR0.4 was more conducive to enhance P solubilization efficacy and available P accumulation. Redundancy analysis indicated Lactobacillus, Spartobacteria and Pseudomonas were key bacteria associated with HCl-Pi especially in AR0.2 and AR0.4. Network analysis showed that increased aeration rate enhanced the connection and function homoplasy among modules and AR0.4 had more orderly community organization for key bacteria to solubilize P in directly and indirectly biotic way. SEM suggested indirectly biotic P-solubilization had more contribution than directly biotic way mainly by phosphate-solubilizing bacteria.

Influence of carbon-to-phosphorus ratios on phosphorus fractions transformation and bacterial community succession in phosphorus-enriched composting.

This study aims to assess the effect of different carbon-to-phosphorus (C:P) ratios on phosphorus (P) fractions transformation, bacterial community succession and microbial P-solubilizing function in kitchen waste composting with rock phosphate (RP) amendment and phosphate-solubilizing bacteria (PSB) inoculation. Results indicated that initial C:P ratio at 50 enhanced organic carbon degradation, available P (AP) accumulation, the amount of PSB and pqqC gene abundance in composting but higher C:P ratio increased microbial biomass phosphorus (MBP) content. Redundancy analysis showed C:P ratios, PSB amount and pqqC gene abundance greatly affected bacterial community diversity and composition. Network analysis indicated that lower C:P ratio enhanced the interaction frequency in core bacterial network for AP transformation. Variance partitioning analysis abiotic factors contributed more to MBP and AP conversion. The study revealed that C:P ratio could directly drive PSB to regulate P fractions and the accumulation of MBP or AP in P-enriched composting.

Integrating 16S rRNA amplicon metagenomics and selective culture for developing thermophilic bacterial inoculants to enhance manure composting.

Composting is an important method for treating and recycling organic waste, and the use of microbial inoculants can increase the efficiency of composting. Herein, we illustrate an approach that integrate 16S rRNA amplicon metagenomics and selective culture of thermophilic bacteria for the development of inoculants to improve manure composting. The 16S rRNA amplicon sequencing analysis revealed that Firmicutes and Actinobacteria were dominant in the composting mixture, and that different microbial hubs succeeded during the thermophilic stage. All isolated thermophilic bacteria were affiliated with the order Bacillales, such as Geobacillus, Bacillus, and Aeribacillus. These isolated thermophilic bacteria were grouped into 11 phylotypes, which shared ＞99% sequence identity to 0.15% to 5.32% of 16S rRNA reads by the amplicon sequencing. Three of these phylotypes transiently enriched during the thermophilic stage. Six thermophilic bacteria were selected from the three phylotypes to obtain seven microbial inoculants. Five out of seven of the microbial inoculants enhanced the thermophilic stage of composting by 16.9% to 52.2%. Three-dimensional excitation emission matrix analysis further revealed that two inoculants (Thermoactinomyces intermedius and Ureibacillus thermophilus) stimulated humification. Additionally, the 16S rRNA amplicon sequencing analysis revealed that inoculation with thermophilic bacteria enhanced the succession of the microbial community during composting. In conclusion, 16S rRNA amplicon metagenomics is a useful tool for the development of microbial inoculants to enhance manure composting.

Impact of inoculation and turning for full-scale composting on core bacterial community and their co-occurrence compared by network analysis.

Window composting with inoculation or frequent turning is a superior way to improve traditional composting efficiency. However, the relationship between the innocent treatment in composting with inoculation or turning and microbial dynamics is unclear. Here, the impact of inoculation and turning for full scale composting on core bacterial community and their co-occurrence network as well as harmless level were compared by network analysis. Results showed that composts with both inoculation and turning had 46% increase of total organic carbon degradation compared to traditional composting and decreased the abundance of potential pathogens. The relative abundance of thermophilic bacteria and Galbibacter, Methylocaldum, Steroidobacter, etc. increased during composting with turning and inoculation. Luteimonas, Sphaerobacter, Turicibacter and Flavobacterium as core bacteria had significant difference between control and composting with enhanced innocent treatment efficiency. Network analysis suggested that turning increased the number of indigenous core bacteria and inoculation enhanced the interaction among key bacterial network.

Give priority to abiotic factor of phosphate additives for pig manure composting to reduce heavy metal risk rather than bacterial contribution.

Phosphate additives especially superphosphate can reduce nitrogen loss, and increase phosphorus availability in composting. This study investigated the changes of different heavy metals fractions and their relationship with bacterial community and abiotic factors during pig manure composting with adding equimolar H3PO4, H2SO4 and K2HPO4. Results showed that both acidic and alkaline labile phosphate increased the potential ecological risk of heavy metals compared to control, but K2HPO4 decreased the accumulation of exchangeable Zn and Mn by 12% and 15% than that with H3PO4 and H2SO4 addition. Network analysis showed that K2HPO4 enhanced the proportion of negative links in bacterial species with heavy metals, but H3PO4 decreased the stability of bacterial network. Redundancy analysis demonstrated that pH was the key factor on metal speciation and risk with phosphate additives than bacterial role. The study presented theoretical basis for additive selection in controlling composting nitrogen fixation and environmental risk.

Dynamics of Diversity and Abundance of Sulfonamide Resistant Bacteria in a Silt Loam Soil Fertilized by Compost.

Although composting is effective in deactivating antibiotic substances in manure, the influence of compost fertilization on the occurrence and dissemination of antibiotic resistance in arable soils remains to be controversial. Herein, the abundance and diversity of two sulfonamide resistance genes (sul1 and sul2) in soil fertilized by compost spiked with two concentrations of sulfadiazine (1 and 10 mg kg-1) were studied intensively by qPCR and high throughput sequencing based on a two-month microcosm experiment. The concentration of sulfadiazine decreased rapidly after spiking from 25% at Day 1 to less than 2.7% at Day 60. Relative abundance of both sul1 and sul2 were significantly higher in soil amended with compost than the non-amended control at Day 1 and slightly decreased with incubation time except for sul2 in the S10 treatment. Soil bacterial communities were transiently shifted by compost fertilization regardless of the presence of sulfadiazine. Relative abundance of genera in three hubs positively interlinked with sul1 and sul2 were significantly higher in compost treated soil than the control at Day 1, 7 and 21, but not at Day 60. High throughput sequencing analyses revealed that most detected (>67% in relative abundance) sul1 and sul2 genotypes sharing >99% similarity with those found in gammaproteobacterial pathogens frequently were commonly present in compost and soil. These results indicated that compost fertilization might increase the abundance rather than diversity of sulfadiazine-resistant populations in soil, which may be facilitated by the presence of sulfadiazine.

Phosphorus excess changes rock phosphate solubilization level and bacterial community mediating phosphorus fractions mobilization during composting.

This study investigated the changes of phosphorus (P) fractions, bacterial community and their response to available P or carbon (C):P during composting with different rock phosphate (RP) addition levels. Results showed that adding RP at 10% or 15% promoted the rise of temperature, maturity and Olsen P accumulation in composting, which had a higher amount of RP solubilization than other groups. Available P changed bacterial composition and decreased diversity in composts. RP solubilization efficiency was negatively correlated to C:P ratio and the highest (22.7%) when 10% RP was added, in which bacterial community changed from "function redundancy" to "intensive P-solubilization". Low C:P ratio (〈300) increased the RP solubilization ratio especially within 135-160. Therefore, this study proposed that adding P-rich substrates to decrease C:P ratio could regulate P-solubilizers' activity for increasing RP solubilization efficiency during composting.

Recycling of nutrients from organic waste by advanced compost technology- A case study.

Composting is widely used as an easily operated and economical method to manage organic wastes. However, the long residence time of composting impedes the recycling nutrients from large amounts of organic wastes produced every day. In this study, the intelligent biodrying + continuous dynamic trough (IB + CDT) was created and used in China's first urban and rural organic waste treatment and utilization demonstration center in Suzhou city. Results showed that IB + CDT composting had higher temperature, more reduction of moisture than windrow composting, enhancing 40% of composting efficiency. Primary fermentation of the IB + CDT composting in the indoor conditions could achieve the harmless treatment (GI > 80%) of compost within 12 days. The IB + CDT composting product enhanced 17% soil organic matter and 11% available nitrogen. The IB + CDT composting mode could earn 57.6 USD/ton by recycling organic waste and producing organic fertilizer, leading to a sustainable and profitable mode.