The binding effects and mechanisms of dissolved organic matter (DOM) on the fate of mercury in sludge anaerobic digestion combined with thermal hydrolysis.

Dissolved organic matter (DOM) plays an important role in regulating the fate of mercury (Hg), e.g., mobility, bioavailability, and toxicity. Clarifying the role of DOM in binding Hg in the treatment processes of sewage sludge is important for relieving Hg contamination risks in land applications. However, the impacts of DOM on Hg binding in sewage sludge are still unclear. In this study, we investigated the evolution of Hg and its speciation in full-scale sludge anaerobic digestion (AD) with thermal hydrolysis. The role of DOM in binding Hg(II) was further analyzed. The results showed that AD with thermal hydrolysis led to an increase in the Hg content in the sludge (from 3.72 ± 0.47 mg/kg to 10.75 ± 0.16 mg/kg) but a decrease in Hg mobility (the mercury sulfide fraction increased from 60.56 % to 79.78 %). Further adsorption experiments revealed that at equivalent DOM concentrations, DOM with a low molecular weight (MW<1 kDa) in activated sludge, DOM with a medium molecular weight (1 kDa 5 kDa) in both anaerobically digested sludge and conditioned sludge showed high binding amounts of Hg(II), with 1372.54, 535.28, 942.09 and 801.51 mg Hg/g DOM, respectively. Parallel factor analysis (PARAFAC) and fluorescence quotient (FQ) results showed that tryptophan-like and tyrosine-like substances had high binding affinities for Hg(II). Furthermore, X-ray photoelectron spectroscopy (XPS) indicated that the reduced organic sulfur contained in the DOM was potentially bound to Hg through the interactions of Hg-S and Hg-O. These results indicated that DOM may play special roles in regulating Hg speciation. The association between DOM and Hg(II), such as the significant positive correlation (p < 0.05) between the dissolution rate of Hg(II) and release of tryptophan-like substances during thermal hydrolysis, suggested the potential way for removing Hg from sludge.

Viral Communities Contribute More to the Lysis of Antibiotic-Resistant Bacteria than the Transduction of Antibiotic Resistance Genes in Anaerobic Digestion Revealed by Metagenomics.

Ecological role of the viral community on the fate of antibiotic resistance genes (ARGs) (reduction vs proliferation) remains unclear in anaerobic digestion (AD). Metagenomics revealed a dominance of Siphoviridae and Podoviridae among 13,895 identified viral operational taxonomic units (vOTUs) within AD, and only 21 of the vOTUs carried ARGs, which only accounted for 0.57 ± 0.43% of AD antibiotic resistome. Conversely, ARGs locating on plasmids and integrative and conjugative elements accounted for above 61.0%, indicating a substantial potential for conjugation in driving horizontal gene transfer of ARGs within AD. Virus-host prediction based on CRISPR spacer, tRNA, and homology matches indicated that most viruses (80.2%) could not infect across genera. Among 480 high-quality metagenome assembly genomes, 95 carried ARGs and were considered as putative antibiotic-resistant bacteria (pARB). Furthermore, lytic phages of 66 pARBs were identified and devoid of ARGs, and virus/host abundance ratios with an average value of 71.7 indicated extensive viral activity and lysis. The infectivity of lytic phage was also elucidated through laboratory experiments concerning changes of the phage-to-host ratio, pH, and temperature. Although metagenomic evidence for dissemination of ARGs by phage transduction was found, the higher proportion of lytic phages infecting pARBs suggested that the viral community played a greater role in reducing ARB numbers than spreading ARGs in AD.

Interpretable Machine Learning and Reactomics Assisted Isotopically Labeled FT-ICR-MS for Exploring the Reactivity and Transformation of Natural Organic Matter during Ultraviolet Photolysis.

Isotopically labeled FT-ICR-MS combined with multiple post-analyses, including interpretable machine learning (IML) and a paired mass distance (PMD) network, was employed to unravel the reactivity and transformation of natural organic matter (NOM) during ultraviolet (UV) irradiation. FT-ICR-MS analysis was used to assign formulas, which were classified on the basis of their molecular compositions and structural categories. Isotope (deuterium, D) labeling was utilized to unequivocally determine the photochemical products and examine the development of OD radical-mediated NOM transformation. With regard to the reactive molecular formulas, CHOS formulas exhibited the highest reactivity (86.5% of precursors disappeared) followed by CHON (53.4%) and CHO (24.6%) formulas. With regard to structural categories, the degree of reactivity decreased in the following order: tannins > condensed aromatics > lignin/CRAMs. The IML algorithm demonstrated that the crucial features governing the reactivity of formulas were the molecular weight, DBE-O, NOSC, and the presence of heteroatoms (i.e., N and S), suggesting that the large and unsaturated compounds containing S and N are more prone to photodegradation. The reactomics approach using the PMD network further indicated that 11 specific molecular formulas in the CHOS and CHO class served as hubs, implying a higher photoreactivity and participation in a range of transformations. The isotope labeling analyses also found that, among the reactions observed, hydroxylation (i.e., +OD) is dominant for lignin/CRAMs and condensed aromatics, and formulas containing ≤10 D atoms were developed. Overall, this study, by adopting rigorous and interpretable techniques, could provide in-depth insights into the molecular-level dynamics of NOM under UV irradiation.

Application of coffee ground-based skeleton builder with FeCl3 for conditions of pre-dewatered sludge toward further deep dewatering.

Skeleton builders are essential for achieving deep sludge dewatering. In this study, a novel spent coffee ground (SCG)-based skeleton builder was developed to attain deep sludge dewatering by combined conditioning with FeCl3, and possible mechanisms were examined. Through different surface analysis techniques, it was demonstrated that at a pyrolysis temperature of 300 °C, the spent coffee ground biochar (SCGB-300) has an intact pore structure, a rigid carbon skeleton, and large oxygen-containing functional groups, making it the best skeleton builder for sludge dewatering. When combined with FeCl3 for conditioning, the structure of SCGB-300 remained intact under high pressure and played important role. The rich porous structure facilitated water drainage. During the sludge conditioning, small amount of positive charge on the surface of SCGB-300 further increased the zeta potential of sludge through charge neutralization. At the same time, the adsorption of SCGB-300 removed viscous hydrophilic substances and further improved the dewatering performance. At an optimum dosage of 6% (dry solid, DS) FeCl3 and 30% SCGB-300 (DS), the moisture content of sludge was reduced from 85.47% to 63.35%, and the dewatering rate was increased from 46.08% to 70.03%. Therefore, SCGB is a promising skeleton builder for sludge conditioning and deep dewatering.

Comprehensive understanding of DOM reactivity in anaerobic fermentation of persulfate-pretreated sewage sludge via FT-ICR mass spectrometry and reactomics analysis.

Understanding the composition and reactivity of dissolved organic matter (DOM) at molecular level is vital for deciphering potential regulators or indicators relating to anaerobic process performance, though it was hardly achieved by traditional analyses. Here, the DOM composition, molecular reactivity and transformation in the enhanced sludge fermentation process were comprehensively elucidated using high-resolution mass spectrometry measurement, and data mining with machine learning and paired mass distance (PMD)-based reactomics. In the fermentation process for dewatered sludge, persulfate (PDS) pretreatment presented its highest performance in improving volatile fatty acids (VFAs) production with the increase from 2,711 mg/L to 3,869 mg/L, whereas its activation in the presence of Fe (as well as the hybrid of Fe and activated carbon) led to the decreased VFAs production performance. In addition to the conventional view of improved decomposition and solubilization of N-containing structures from sludge under the sole PDS pretreatment, the improved VFAs production was associated with the alternation of DOM molecular compositions such as humification generating molecules with high O/C, N/C, S/C and aromatic index (AImod). Machine learning was capable of predicting the DOM reactivity classes with 74-76 % accuracy and found that these molecular parameters in addition to nominal oxidation state of carbon (NOSC) were among the most important variables determining the generation or disappearance of bio-resistant molecules in the PDS pretreatment. The constructed PMD-based network suggested that highly connected molecular network with long path length and high diameter was in favor of VFAs production. Especially, -NH related transformation was found to be active under the enhanced fermentation process. Moreover, network topology analysis revealed that CHONS compounds (e.g., C13H27O8N1S1) can be the keystone molecules, suggesting that the presence of sulfur related molecules (e.g., cysteine-like compounds) should be paid more attention as potential regulators or indicators for controlling sludge fermentation performance. This study also proposed the non-targeted DOM molecular analysis and downstream data mining for extending our understanding of DOM transformation at molecular level.

Understanding of mercury and methylmercury transformation in sludge composting by metagenomic analysis.

Municipal sewage especially the produced sewage sludge is a significant source releasing mercury (Hg) to the environment. However, the Hg speciation especially methylmercury (MeHg) transformation in sewage sludge treatment process remains poorly understood. This study investigated the transformation of Hg speciation especially MeHg in sludge composting. The distribution of Hg transformation related gene pairs hgcAB and merAB, and their putative microbial hosts were comprehensively analyzed. Both Hg (from 3.16±0.22 mg/kg to 3.20±0.19 mg/kg) and MeHg content (from 4.77±0.64 ng/g to 4.36±0.37 ng/g) were not obviously changed before and after composting, but about 19.69% of Hg and 27.36% of MeHg were lost according to mass balance calculation. The metagenomic analysis further revealed that anaerobes (Desulfobacterota and Euryarchaeota) were the mainly putative Hg methylators especially carrying high abundance of hgcA gene in the initial periods of composting. Among the 151 reconstructed metagenome-assembled genomes (MAGs), only 4 hgcA gene carriers (Myxococcota, Firmicutes, Cyclobacteriaceae, and Methanothermobacter) and 16 merB gene carriers were identified. But almost all of the MAGs carried hgcB gene and merA gene. The merA gene was widely distributed in genomes, which indicated the widespread functionality of microbes for reducing Hg(II) to Hg(0). The hgcA carrying microbes tends to present the similar metabolic pathways including methanogenesis and sulfur metabolism. Besides, both the irregular distribution of hgcA in various species (including Actinobacteria, Archaea, Bacteroidetes, Desulfobacterota, Euryarchaeota, and Nitrospirae, etc.) and opposite evolution trends between hgcA gene abundance and its host genome abundance can be an indication of horizontal gene transfer or gene deletions of hgcA during composting. Our findings thus revealed that sludge composting is not only a hotspot for Hg speciation transformation, but also a potential hotspot for MeHg transformation.

Removal of methylmercury and its potential relationship to microbiota in sludge anaerobic digestion under thermal hydrolysis.

Reducing health risk of mercury (Hg)/methylmercury (MeHg) in sewage sludge is vital to its land application. This study revealed that thermal hydrolysis reduced MeHg content both during pretreatment process and subsequent anaerobic digestion (AD), which resulted in decrease of MeHg content from 4.24 ng/g to 0.95 ng/g after thermal hydrolysis (150 ℃) and further decreased to 0.39 ng/g after AD. Notably, thermal hydrolysis at high temperature (120 ℃ and 150 ℃) promoted both Hg methylation and MeHg demethylation rather than the control or at low temperature (100 ℃). Hg methylation dominated in hydrolysis and acidogenesis stage, whereas MeHg demethylation dominated in methanogenesis stage. Though abundance of related genes (HgcA and merA) was dramatically reduced, Ruminococcaceae, Peptococcaceae, and Lachnospiraceae were potentially Hg methylators in hydrolysis and acidogenesis stage. Whereas, MeHg demethylation dominated in the late period of AD due to the improved syntrophic methanogenesis and possibly reduced Hg2+ biodegradability by precipitation.

Effect of rainfall characteristics on the sewer sediment, hydrograph, and pollutant discharge of combined sewer overflow.

Significant management needs raised in urban sewer system to facilitate urban resilience to rainstorm. The work investigated the effects of temporal evolution of rainfall on hydrograph and pollutant discharge of CSO over an intensive observation period of 12 months, with special attention to differences in temporal scale for supporting management decision making. The characteristics of rainfall in different temporal scales helped overflow-risk identification and assessment. Prolonged dry seasons over 112 days in the CSO monitored year 2018 increased the sediment buildup in the pipes. The built sediment was mostly flushed out to overflow (and the treatment facility) by initial rainfall during 47 h. Following CSO hydraulics and pollutant discharge follows initial peak patterns which responded to rainfall patterns. Results of Redundancy analysis and network analysis showed that the combined effects of rainfall, urbanization, and sediments as "CSO troika" are the driving forces for CSO pollutants in the long-term. The improved characterization of CSO events and the associated pollutants has refined our understanding of how overflow hydrograph and pollutant discharge responds to rainfall temporally, which methodology supported decision making in the combining source/process control with terminal management for facilizing urban resilience.

Delivery and effects of proton pump inhibitor on anaerobic digestion of food and kitchen waste under ammonia stress.

Ammonia stress changes microbial metabolism of anaerobic digestion and decreases methane yield, where proton pump overactivated by free ammonia suggested to be the centre of the metabolism changes in anaerobic digestion under ammonia stress. The work demonstrated that proton pump inhibitor (PPI) could alleviate the overactivated proton pump and mitigate ammonia inhibition. Its impacts on iron transporter, substrates uptake, and energy conservation were investigated in anaerobic digestion treating food and kitchen waste. The PPI formed a stimuli-responsive drug delivery system driven by pH for the more inhibited microbe (p < 0.01), confirmed by FE-SEM/EDS and high throughput sequencing, implying the PPI was activated at inhibited microbe more than mixed liquor. Consistent microbial population increase observed in syntrophs and methanogens, who utilized the substrates for high yielding pathway and facilitated the energy sharing by direct interspecies electron transfer. These results demonstrated PPI could recovery methane production and could mitigate fatty-acid accumulation under high ammonia stress by delivery and activation in acetoclastic methanogen.

Fate of mercury and methylmercury in full-scale sludge anaerobic digestion combined with thermal hydrolysis.

Methylmercury (MeHg) is one of the highly toxic and bio-accumulated forms of mercury. Its presence in wastewater treatment processes has been evidenced in recent studies. Considering its enrichment in sewage sludge and the ecological risk associated with its land application, this study investigated the fate of mercury and MeHg in full-scale anaerobic digestion combined with Cambi thermal hydrolysis based on one-year sampling. Results showed that the advanced anaerobic digestion could increase the total mercury (THg) content from 4.35 ± 0.43 mg/kg in raw sludge to 6.37 ± 1.05 mg/kg in digested sludge, and the MeHg content decreased from 1.61 to 8.94 ng/g in raw sludge to 0.21-2.03 ng/g after anaerobic digestion. The demethylation of MeHg was dominant in both thermal hydrolysis and anaerobic digestion; it was mostly derived from the physico-chemical impacts such as chemical decomposition in thermal hydrolysis and precipitation in anaerobic digestion. Although the reported microbial methylators, such as Methanosarcina and Clostridia, were dominant in anaerobic digestion, the relative abundances of hgcA and merA were relatively low and did not correlate with the MeHg profiles. Thus, microbial methylation or demethylation seems negligible in terms of MeHg transformation.

Effect of proton pump inhibitor on microbial community, function, and kinetics in anaerobic digestion with ammonia stress.

The proton pump is a convincing mechanism for ammonia inhibition in anaerobic digestion, which explained how the ammonia accumulated intercellularly due to diffusion of free ammonia. Proton pump inhibitor (PPI) was dosed for mitigating the accumulation in anaerobic digestion with ammonia stress, with respect to kinetics. Results show PPI inhibited ß-oxidation of fatty acids by targeting ATPase in anaerobic digestion with ammonia stress. Alternatively, PPI stimulated syntrophic acetate oxidization. Random forest located key genera as syntrophic consortia. Methane increased 18.72 ± 7.39% with 20 mg/L PPI at the first peak, consistent with microbial results. The deterministic Gompertz kinetics and stochastic Gaussian processes contributed 97.63 ± 8.93% and 2.37 ± 8.93% in accumulated methane production, respectively. Thus, the use of PPI for anaerobic digestion allowed mitigate ammonia inhibition based on the mechanism of proton pump, facilitate intercellularly ammonia accumulation, stimulate syntrophic consortia, and eliminate uncertainty of process failure, which resulted in efficient methane production under ammonia stress.

[Effect on Ammonia Inhibition Mitigation in the Anaerobic Digestion Process with Zero-Valent Iron].

Ammonia inhibition is an important factor impacting methane production efficiency during the anaerobic digestion of high-solid organic wastes. This study investigated the effect of micro-sized zero-valent iron (m-ZVI) on the anaerobic digestion of excess sewage sludge and thermal hydrolyzed sludge using batch mode experiments. The effect of m-ZVI on ammonia inhibition mitigation was also studied. Results showed that the kinetic characteristics of the methane production rate, lag phase, and methane production potential of the anaerobic digestion of excess sludge and thermal hydrolyzed sludge were not impacted by the addition of m-ZVI at a dosage of 4 g·L-1 and 10 g·L-1. However, during the inhibited anaerobic digestion process with a high ammonia concentration, the addition of 4 g·L-1 and 10 g·L-1 of m-ZVI was able to shorten the lag phase from 18.61 d (the control) to 17.22 d and 16.18 d, respectively. Moreover, the maximum methane production rate (based on the VS) increased from 6.34 mL·(d·g)-1(the control) to 7.84 mL·(d·g)-1 (4 g·L-1 m-ZVI) and 7.39 mL·(d·g)-1 (10 g·L-1 m-ZVI). The pH buffer system was not influenced by the chemical reaction of m-ZVI in the anaerobic digestion, although the relative abundance of the dominant methanogenic archaea (Methanosarcina) improved greatly from 30.71% (the control) to 53.50% (4 g·L-1 m-ZVI) and 60.30% (10 g·L-1 m-ZVI) at 27 d. This study proved that m-ZVI was incapable of improving the methane production potential of sewage sludge, while the mitigation of ammonia inhibition during anaerobic digestion was enhanced by the stimulating effect on methanogenic archaea.

Relieving ammonia inhibition by zero-valent iron (ZVI) dosing to enhance methanogenesis in the high solid anaerobic digestion of swine manure.

Relieving from ammonia inhibition and enhancing the utilization of thermodynamically unfavorable propionate are crucial for methane harvest in the high solid anaerobic digestion (HSAD) of ammonia-rich swine manure. In this study, the potential of dosing zero-valent iron (ZVI, 150 um) for enhancing the methanogenesis to resist total ammonia (TAN) over 5.0 g-N·L-1 was investigated via batch experiments under mesophilic condition. The cumulative methane production was enhanced by 22.2% at ≥160 mM ZVI dosage and the HSAD duration was further shortened by 50.6% at ≥320 mM ZVI dosage. The enhanced methanogenesis was mainly resulted from the full utilization of propionate and the accelerated collapse of posterior-biodegradable organics which might be driven by ZVI. Results of microbial community and qPCR (mcrA) showed that ZVI might trigger the blooming of Methanosarcina (from 27.9% to 78.3%) and Syntrophomonas (0.5% to 3.7%) and attribute to their possible direct interspecies electron transfer (DIET) to enhance propionate utilization. Besides, the main methanogenesis might remain in the effective aceticlastic pathway even under free ammonia (FAN) almost 1.0 g-N·L-1 because syntrophic acetate oxidizing bacteria (SAOB) decreased to almost none at 320 mM ZVI dosage. Dosing ZVI could relieve HSAD from TAN inhibition and more dosage was required to resist FAN inhibition.

N2O emission factors of full-scale animal manure windrow composting in cold and warm seasons.

Emission of nitrous oxide (N2O) during animal manure composting is of great concern, and its emission factor (EF) is important for greenhouse gas emission inventory, while the EF is still uncertain due to limited on-site full-scale observations worldwide. In this study, N2O emissions were monitored during different seasons in a full-scale swine manure windrow composting with pile volume of about 76.5 m3. The results showed that the maximum N2O flux during the cold season (CS) was 23 times higher than during the warm season (WS), significant differences in the contribution to direct N2O emissions were observed in three composting stages, and shaded-side N2O emission was higher than sunny-side emission. The direct N2O emission factors of animal manure composting were 0.0046, 0.0002 kg N2O-N/kgTN (dry weight) in the CS and WS, respectively. Scenario analysis results showed that windrow composting is a suitable manure management that emits less N2O than solid storage.

[Effect of Sulfate on the Migration and Transformation of Methylmercury in Advanced Anaerobic Digestion of Sludge].

To study the migration and transformation of methylmercury during advanced anaerobic digestion of sludge and the role of sulfate, this study investigated the migration and transformation of methylmercury during different stages of sludge anaerobic digestion (AD) with thermal hydrolysis pretreatment and under different dosages of sulfate addition. The results showed that mercury methylation occurred in the initial stage of AD (Day 1-3), the ratio of methylmercury to total mercury increased from 0.024% (range of 0.019%-0.033%) to 0.038% (range of 0.030%-0.048%), and the net increment of methylmercury increased by 3.97, 6.09, 0.17, 3.71, and 1.66 times, respectively. In the following Day 3-5, the demethylation process occurred with the net yield of methylmercury decreased by 71.25% (ranging from 67.42% to 75.10%). Sulfate inhibited the methylation of mercury in the initial stage of AD, but had little effect on it in the late stage. This was related to the reduction of the bioavailability of neutral mercury complexes by charged groups of HgHS22- and HgS22-, as well as the immobilization of iron sulfide and mercury sulfide on S2- and bioavailable mercury. Redundancy analysis (RDA) showed that mercury methylation was affected by several factors:organic substances such as propionic acid, isobutyric acid, isovaleric acid, and Fe may promote mercury methylation, whereas protein and higher pH may be inhibitors of mercury methylation in AD of sludge.

Effect of zero-valent iron combined with carbon-based materials on the mitigation of ammonia inhibition during anaerobic digestion.

Ammonia inhibition is a prominent problem for anaerobic digestion (AD) of nitrogen-rich organic wastes. This study evaluated the effect of zero valent iron (ZVI) and its hybrid with activated carbon (AC), graphite and Fe-C material on the mitigation of ammonia inhibition under ammonia concentration over 5 g/L, according to the batch mode experiments. Results showed that ZVI (4 g/L) and its hybrid with carbon-based material preserving methane production from ammonia inhibition, with kinetics of shortening lag phase from 4.77 d to 2.62 d or even below 2 d with carbon-based material. ZVI preserved methane production with the enrichment of Methanosarcina (the relative abundance was over 80%), which was mostly derived from the activating hydrogenotrophic methanogenesis through the enhanced DIET but not the changes of ORP and FAN.

The performance evaluation and kinetics response of advanced anaerobic digestion for sewage sludge under different SRT during semi-continuous operation.

Sludge retention time (SRT) is vital for advanced anaerobic digestion (AD) to realize energy self-sufficient. However, the criteria on reasonable SRT has not been fully understood. This study investigated the performance and kinetics response of AD under different SRT in semi-continuous AD with microwave (MW) pretreatment, according to the long-term operation and methane production during one feeding interval. Results showed that modified Gompertz model better described the kinetics than first-order model. At short SRT (15 d), pretreatment coupled with two-stage AD preserved methane production with the high attainable methane potential (B0) of 257.98 mL/g VS and hydrolysis rate constant (khyd) of 0.075 h-1. But the acceptable decrease of methane production rate seems to be unavoidable, which was possibly derived from the evolution of methanogenesis pathway. This study emphasized the importance of improved methane production rate in semi-continuous AD under short SRT rather than methane production potential obtained from batch experiment.

[Enhancement for Anaerobic Digestion of Waste Activated Sludge Based on Microwave Pretreatment Combined with Zero Valent Iron].

In this study, we investigated the enhanced performance after addition of zero valent iron (ZVI) under different dosages (low range of 5.19-41.51 g·kg-1 TS and high range of 83.35-853.46 g·kg-1 TS), combined with microwave (MW) pretreatment for anaerobic digestion (AD) of waste activated sludge (WAS). The results demonstrated that the methane production potential of WAS could be increased by 17%-24% with the addition of ZVI combined with MW pretreatment, and especially the methane production rate was enhanced in the initial days (1-4d). ZVI addition could further improve the enhanced performance of AD under MW pretreatment. Compared with the performance of AD with only MW pretreatment, the methane production potential was increased by 7.42%, and methane production flow rate at 2 d was increased by 11.02% with 31.13 g·kg-1 TS of ZVI addition. However, the higher dosage of added ZVI did not show further enhanced performance. It was concluded that ZVI addition promoted the release of dissolved organics at the initial stage of AD. For instance, soluble proteins were increased by 21.16% with the ZVI addition of 31.13 g·kg-1 TS compared with pretreated WAS without ZVI addition. Furthermore, ZVI addition accelerated the degradation of acetic acid, iso-butyric acid, and iso-valeric acid, and led to a significant reduction of orthophosphate and sulfate in the supernatant of the digested sludge. The concentration of iron in the supernatant decreased even with a high dosage of ZVI. Thus, the formation of precipitate that occurred due to reactions between iron and orthophosphate or sulfate, may be the main reason for the lack of enhanced performance even with high dosage of ZVI addition.

The role of substrate types and substrate microbial community on the fate of antibiotic resistance genes during anaerobic digestion.

Anaerobic digestion (AD) is regarded as a promising technology in energy recovery and the spread mitigation of antibiotic resistance. However, the performance of AD is dependent on various factors, and substrate type is one of the most important. In this study, the fate of antibiotic resistance genes (ARGs) response to the substrate types was investigated, and three typical environmental reservoirs of ARGs (pig manure, chicken manure and sewage sludge) were selected. The role of substrate microbial community on the fate of ARGs was clarified through the comparison between the AD of the substrates with and without a prior autoclave-disinfected step. Results showed that substrate types significantly influenced the fate of ARGs, while the influence from the substrate microbial community was limited. The concentration of antibiotics, the horizontal gene transfer reflected by intI1 and co-selection from heavy metals reflected by metal resistance genes (MRGs) were all reduced effectively. Microbial community varied from substrate types and dominated the ARGs fate concerning the standardized total effects through the mantel test and SEM analysis. The fate of tetX, ermF, tetM and ermB was mainly determined by the physicochemical parameters and the phyla of Firmicutes and Bacteroides. The phyla of Actinobacteria, pcoA and czcA contributed most to the reduction of blaTEM and mcr-1, and the phyla of Proteobacteria, Chloroflexi, Synergistetes, Euryarchaeote, intI1 and merA correlated significantly with the fate of blaCTX-M, ereA, tetG and sulI. This study highlighted the importance of substrate types when considering the fate of ARGs during AD.

Fate of antibiotic resistance genes during anaerobic digestion of sewage sludge: Role of solids retention times in different configurations.

In this study, three anaerobic digestion experiments were established to investigate the effects of solids retention times (SRT) on the fate of antibiotic resistance genes (ARGs) including anaerobic digestion of sewage sludge (CK), one-stage anaerobic digestion of microwave pretreatment sludge (MW) and two-stage anaerobic digestion of microwave pretreatment sludge (Acid stage and CH4 stage). The response of ARGs to the SRT varied significantly from ARG types and reactor configurations. Shorter SRT could avail the ARGs reduction for CK and two-stage digestion, while MW need longer SRT for the ARGs reduction. Concerning the variance of microbial community caused by reactor configurations, the role of SRT was limited. The partial redundancy analysis and structural equation models analysis indicated that the role of SRT on the ARGs fate could be attributed the most to the co-selection from heavy metals.