Metagenomic analysis on resistance genes in water and microplastics from a mariculture system

• Total 174 subtypes of ARGs were detected by metagenomic analysis. • Chloramphenicol resistance genes were the dominant ARGs in water and microplastics. • The abundances of MRGs were much higher than those of ARGs. • Proteobacteria, Bacteroidetes, and Actinobacteria were the dominant phylum. • Microplastics in mariculture system could enrich most of MRGs and some ARGs. Microplastics existing widely in different matrices have been regarded as a reservoir for emerging contaminants. Mariculture systems have been observed to host microplastics and antibiotic resistance genes (ARGs). However, more information on proliferation of ARGs and metal resistance genes (MRGs) in mariculture system at the presence of microplastics is needed. This study used metagenomic analysis to investigate the distribution of ARGs and MRGs in water and microplastics of a typical mariculture pond. Total 18 types including 174 subtypes of ARGs were detected with the total relative abundances of 1.22/1.25 copies per 16S rRNA copy for microplastics/water. Chloramphenicol resistance genes were the dominant ARGs with the abundance of 0.35/0.42 copies per 16S rRNA copy for microplastics/water. Intergron intI1 was dominant gene among 6 detected mobile genetic elements (MGEs) with the abundance of 75.46/68.70 copies per 16S rRNA copy for water/microplastics. Total 9 types including 46 subtypes of MRGs were detected with total abundance of 5.02 × 10²/6.39 × 10² copies per 16S rRNA copy for water/ microplastics while genes resistant to copper and iron served as the dominant MRGs. Proteobacteria, Bacteroidetes, and Actinobacteria accounted for 84.2%/89.5% of total microbial community. ARGs with relatively high abundance were significantly positively related to major genera, MGEs, and MRGs. Microplastics in mariculture system could enrich most of MRGs and some ARGs to serve as potential reservoir for these pollutants. The findings of this study will provide important information on resistance gene pollution at presence of microplastics in the mariculture system for further proposing suitable strategy of environmental management.     

Distribution of antibiotic resistance genes and their association with bacteria and viruses in decentralized sewage treatment facilities

• Distribution of ARGs in decentralized sewage facilities were investigated. • Bacitracin-ARGs were most predominant ARGs in rural wastewater. • ARGs were identified in bacterial and viral community. • ARGs of rpoB, drfE, gyrA and parC were both correlated with bacteria and phages. • More attention should be paid to the risk of spreading ARG by phages. The distribution of antibiotic resistance genes (ARGs) has been intensively studied in large-scale wastewater treatment plants and livestock sources. However, small-scale decentralized sewage treatment facilities must also be explored due to their possible direct exposure to residents. In this study, six wastewater treatment facilities in developed rural areas in eastern China were investigated to understand their risks of spreading ARGs. Using metagenomics and network analysis tools, ARGs and bacterial and viral communities were identified in the influent (INF) and effluent (EFF) samples. The dominant ARGs belonged to the bacitracin class, which are different from most of municipal wastewater treatment plants (WWTPs). The dominant hosts of ARGs are Acidovorax in bacterial communities and Prymnesiovirus in viral communities. Furthermore, a positive relationship was found between ARGs and phages. The ARGs significantly correlated with phages were all hosted by specific genera of bacteria, indicating that phages had contributed to the ARG’s proliferation in sewage treatment facilities. Paying significant concern on the possible enhanced risks caused by bacteria, viruses and their related ARGs in decentralized sewage treatment facilities is necessary.     

Antibiotic resistance genes in manure-amended paddy soils across eastern China: Occurrence and influencing factors

• Manure fertilization resulted in antibiotic residues and increased metal contents. • The tet and sul genes were significantly enhanced with manure fertilization. • Soil physicochemical properties contributed to 12% of the variations in ARGs. • Soil metals and antibiotics co-select for ARGs. Pig manure, rich in antibiotics and metals, is widely applied in paddy fields as a soil conditioner, triggering the proliferation of antibiotic resistance genes (ARGs) in soil. However, comprehensive studies on the effects of manure fertilization on the abundance of ARGs and their influencing factors are still insufficient. Here, pig manure and manure-amended and inorganic-amended soils were collected from 11 rice-cropping regions in eastern China, and the accumulation of antibiotics, metals, and ARGs was assessed simultaneously. The results showed that manure fertilization led to antibiotic residues and increased the metal content (i.e., Zn, Cu, Ni, and Cr). Tetracycline and sulfonamide resistance genes (tetM, tetO, sul1, and sul2) were also significantly enhanced with manure fertilization. According to variance partitioning analysis, the most important factors that individually influenced ARGs were soil physicochemical properties, accounting for 12% of the variation. Significant correlations between soil nutrients and ARGs indicated that manure application enhanced the growth of resistant microorganisms by supplying more nutrients. Metals and antibiotics contributed 9% and 5% to the variations in ARGs, respectively. Their co-occurrence also increased the enrichment of ARGs, as their interactions accounted for 2% of the variation in ARGs. Interestingly, Cu was significantly related to most ARGs in the soil (r = 0.26–0.52, p<0.05). Sulfapyridine was significantly related to sul2, and tetracycline resistance genes were positively related to doxycycline. This study highlighted the risks of antibiotic and ARG accumulation with manure fertilization and shed light on the essential influencing factors of ARGs in paddy soils.     

Effect of the ultraviolet/chlorine process on microbial community structure,typical pathogens,and antibiotic resistance genes in reclaimed water

• UV/chlorine can effectively remove VBNC pathogens, ARGs and MGEs in reclaimed water. • Microbial community was changed with reduced diversity during UV/chlorine process. • CRBs-carried MGEswere the predominant groups during UV/chlorine process. • No direct co-selection strategy was shared between UV/chlorine and resistome. Urban wastewater contains a wide range of pathogens and antibiotic resistance genes (ARGs), which are a serious concern if reusing treated wastewater. However, few studies have explored the microbial communities in reclaimed water using ultraviolet (UV)/chlorine treatment and assessed the changes of the resistome. This study investigated the occurrence of typical pathogens, ARGs, and bacterial communities in UV/chlorine-treated reclaimed water samples. The numbers of culturable and viable but non-culturable pathogens were effectively reduced to 0 CFU/mL within 1–10 and 10–30 min after UV/chlorine treatment, respectively. Meanwhile, the physicochemical indices of water quality were not affected. UV/chlorine treatment could significantly change the bacterial community structure of reclaimed water, showing a decrease in bacterial abundance and diversity. Chlorine-resistant Acinetobacter and Mycobacterium were the dominant bacterial genera (>50%) after UV/chlorine treatment. Moreover, the number of ARGs and mobile genetic elements (MGEs) decreased with an increase in UV/chlorine exposure. However, eight ARGs and three MGEs were consistently detected in more than three seasons, making these major concerns because of their potential role in the persistence and dissemination of antibiotic resistance. Overall, the results of this study suggest that UV/chlorine treatment can potentially improve the microbiological safety of reclaimed water. And more attention should be paid to the pathogens that are both chlorine-resistant and carry MGEs because of their potential for resistance transmission.     

Degradation of polyacrylamide (PAM) and methane production by mesophilic and thermophilic anaerobic digestion: Effect of temperature and concentration

• PAM degradation in thermophilic AD in comparison with mesophilic AD. • PAM degradation and its impact on thermophilic and mesophilic AD. • Enhanced methane yield in presence of PAM during thermophilic and mesophilic AD. • PAM degradation and microbial community analysis in thermophilic and mesophilic AD. Polyacrylamide (PAM) is generally employed in wastewater treatment processes such as sludge dewatering and therefore exists in the sludge. Furthermore, it degrades slowly and can deteriorate methane yield during anaerobic digestion (AD). The impact or fate of PAM in AD under thermophilic conditions is still unclear. This study mainly focuses on PAM degradation and enhanced methane production from PAM-added sludge during 15 days of thermophilic (55°C) AD compared to mesophilic (35°C) AD. Sludge and PAM dose from 10 to 50 g/kg TSS were used. The results showed that PAM degraded by 76% to 78% with acrylamide (AM) content of 0.2 to 3.3 mg/L in thermophilic AD. However, it degraded only 27% to 30% with AM content of 0.5 to 7.2 mg/L in mesophilic AD. The methane yield was almost 230 to 238.4 mL/g VSS on the 8th day in thermophilic AD but was 115.2 to 128.6 mL/g VSS in mesophilic AD. Mechanism investigation revealed that thermophilic AD with continuous stirring not only enhanced PAM degradation but also boosted the organics release from the sludge with added PAM and gave higher methane yield than mesophilic AD.     

Assessment of antibiotic resistance genes in dialysis water treatment processes

• Quantitative global ARGs profile in dialysis water was investigated. • Totally 35 ARGs were found in the dialysis treatment train. • 29 ARGs (highest) were found in carbon filtration effluent. • erm and mtrD-02 occurred in the final effluent. • The effluent was associated with health risks even after RO treatment. Dialysis water is directly related to the safety of hemodialysis patients, thus its quality is generally ensured by a stepwise water purification cascade. To study the effect of water treatment on the presence of antibiotic resistance genes (ARGs) in dialysis water, this study used propidium monoazide (PMA) in conjunction with high throughput quantitative PCR to analyze the diversity and abundance of ARGs found in viable bacteria from water having undergone various water treatment processes. The results indicated the presence of 35 ARGs in the effluents from the different water treatment steps. Twenty-nine ARGs were found in viable bacteria from the effluent following carbon filtration, the highest among all of the treatment processes, and at 6.96 Log (copies/L) the absolute abundance of the cphA gene was the highest. Two resistance genes, erm (36) and mtrD-02, which belong to the resistance categories macrolides-lincosamides-streptogramin B (MLSB) and other/efflux pump, respectively, were detected in the effluent following reverse osmosis treatment. Both of these genes have demonstrated the potential for horizontal gene transfer. These results indicated that the treated effluent from reverse osmosis, the final treatment step in dialysis-water production, was associated with potential health risks.     

Biofiltration and disinfection codetermine the bacterial antibiotic resistome in drinking water: A review and meta-analysis

• Published data was used to analyze the fate of ARGs in water treatment. • Biomass removal leads to the reduction in absolute abundance of ARGs. • Mechanism that filter biofilm maintain ARB/ARGs was summarized. • Potential BAR risks caused by biofiltration and chlorination were proposed. The bacterial antibiotic resistome (BAR) is one of the most serious contemporary medical challenges. The BAR problem in drinking water is receiving growing attention. In this study, we focused on the distribution, changes, and health risks of the BAR throughout the drinking water treatment system. We extracted the antibiotic resistance gene (ARG) data from recent publications and analyzed ARG profiles based on diversity, absolute abundance, and relative abundance. The absolute abundance of ARG was found to decrease with water treatment processes and was positively correlated with the abundance of 16S rRNA (r² = 0.963, p<0.001), indicating that the reduction of ARG concentration was accompanied by decreasing biomass. Among treatment processes, biofiltration and chlorination were discovered to play important roles in shaping the bacterial antibiotic resistome. Chlorination exhibited positive effects in controlling the diversity of ARG, while biofiltration, especially granular activated carbon filtration, increased the diversity of ARG. Both biofiltration and chlorination altered the structure of the resistome by affecting relative ARG abundance. In addition, we analyzed the mechanism behind the impact of biofiltration and chlorination on the bacterial antibiotic resistome. By intercepting influent ARG-carrying bacteria, biofilters can enrich various ARGs and maintain ARGs in biofilm. Chlorination further selects bacteria co-resistant to chlorine and antibiotics. Finally, we proposed the BAR health risks caused by biofiltration and chlorination in water treatment. To reduce potential BAR risk in drinking water, membrane filtration technology and water boiling are recommended at the point of use.     

Environmental effects and risk control of antibiotic resistance genes in the organic solid waste aerobic composting system: A review

• ARGs were detected in livestock manure, sludge, food waste and fermentation dregs. • The succession of microbial community is an important factor affecting ARGs. • Horizontal transfer mechanism of ARGs during composting should be further studied. Antibiotic resistance genes (ARGs) have been diffusely detected in several kinds of organic solid waste, such as livestock manure, sludge, antibiotic fermentation residues, and food waste, thus attracting great attention. Aerobic composting, which is an effective, harmless treatment method for organic solid waste to promote recycling, has been identified to also aid in ARG reduction. However, the effect of composting in removing ARGs from organic solid waste has recently become controversial. Thus, this article summarizes and reviews the research on ARGs in relation to composting in the past 5 years. ARGs in organic solid waste could spread in different environmental media, including soil and the atmosphere, which could widen environmental risks. However, the conventional composting technology had limited effect on ARGs removal from organic solid waste. Improved composting processes, such as hyperthermophilic temperature composting, could effectively remove ARGs, and the HGT of ARGs and the microbial communities are identified as vital influencing factors. Currently, during the composting process, ARGs were mainly affected by three response pathways, (I) “Microenvironment-ARGs”; (II) “Microenvironment-microorganisms-ARGs”; (III) “Microorganisms-horizontal gene transfer-ARGs”, respectively. Response pathway II had been studied the most which was believed that microbial community was an important factor affecting ARGs. In response pathway III, mainly believed that MGEs played an important role and paid less attention to eARGs. Further research on the role and impact of eARGs in ARGs may be considered in the future. It aims to provide support for further research on environmental risk control of ARGs in organic solid waste.     

Acid Orange 7 degradation using methane as the sole carbon source and electron donor

• AO7 degradation was coupled with anaerobic methane oxidation. • Higher concentration of AO7 inhibited the degradation. • The maximum removal rate of AO7 reached 280 mg/(L·d) in HfMBR. • ANME-2d dominated the microbial community in both batch reactor and HfMBR. • ANME-2d alone or synergistic with the partner bacteria played a significant role. Azo dyes are widely applied in the textile industry but are not entirely consumed during the dyeing process and can thus be discharged to the environment in wastewater. However, azo dyes can be degraded using various electron donors, and in this paper, Acid Orange 7 (AO7) degradation performance is investigated using methane (CH₄) as the sole electron donor. Methane has multiple sources and is readily available and inexpensive. Experiments using ¹³C-labeled isotopes showed that AO7 degradation was coupled with anaerobic oxidation of methane (AOM) and, subsequently, affected by the initial concentrations of AO7. Higher concentrations of AO7 could inhibit the activity of microorganisms, which was confirmed by the long-term performance of AO7 degradation, with maximum removal rates of 8.94 mg/(L·d) in a batch reactor and 280 mg/(L·d) in a hollow fiber membrane bioreactor (HfMBR). High-throughput sequencing using 16S rRNA genes showed that Candidatus Methanoperedens, affiliated to ANME-2d, dominated the microbial community in the batch reactor and HfMBR. Additionally, the relative abundance of Proteobacteria bacteria (Phenylobacterium, Pseudomonas, and Geothermobacter) improved after AO7 degradation. This outcome suggested that ANME-2d alone, or acting synergistically with partner bacteria, played a key role in the process of AO7 degradation coupled with AOM.     

Influences and mechanisms of nanofullerene on the horizontal transfer of plasmid-encoded antibiotic resistance genes between E. coli strains

• Sub-inhibitory levels of nC₆₀ promote conjugative transfer of ARGs. • nC₆₀ can induce ROS generation, oxidative stress and SOS response. • nC₆₀ can increase cell membrane permeability and alter gene expression. • Results provide evidence of nC₆₀ promoting antibiotic resistance dissemination. The spread and development of antibiotic resistance globally have led to severe public health problems. It has been shown that some non-antibiotic substances can also promote the diffusion and spread of antibiotic resistance genes (ARGs). Nanofullerene (nC₆₀) is a type of nanomaterial widely used around the world, and some studies have discovered both the biological toxicity and environmental toxicity of nC₆₀. In this study, cellular and molecular biology techniques were employed to investigate the influences of nC₆₀ at sub-minimum inhibitory concentrations (sub-MICs) on the conjugation of ARGs between the E. coli strains. Compared with the control group, nC₆₀ significantly increased the conjugation rates of ARGs by 1.32‒10.82 folds within the concentration range of 7.03‒1800 mg/L. This study further explored the mechanism of this phenomenon, finding that sub-MICs of nC₆₀ could induce the production of reactive oxygen species (ROS), trigger SOS-response and oxidative stress, affect the expression of outer membrane proteins (OMPs) genes, increase membrane permeability, and thus promote the occurrence of conjugation. This research enriches our understanding of the environmental toxicity of nC₆₀, raises our risk awareness toward nC₆₀, and may promote the more rational employment of nC₆₀ materials.     

Fate and risk assessment of emerging contaminants in reclaimed water production processes

• PPCPs had the highest removal efficiency in A²O combined with MBR process (86.8%). • ARGs and OPFRs were challenging to remove (6.50% and 31.0%, respectively). • Octocrylene and tris(2-ethylhexyl) phosphate posed high risks to aquatic organisms. • Meta-analysis was used to compare the ECs removal in wastewater treatment. • Membrane treatment technology is the most promising treatment for ECs removal. Reclaimed water has been widely applied in irrigation and industrial production. Revealing the behavior of emerging contaminants in the production process of reclaimed water is the first prerequisite for developing relevant water quality standards. This study investigated 43 emerging contaminants, including 22 pharmaceuticals and personal care products (PPCPs), 11 organophosphorus flame retardants (OPFRs), and 10 antibiotic resistance genes (ARGs) in 3 reclaimed wastewater treatment plants (RWTPs) in Beijing. The composition profiles and removal efficiencies of these contaminants in RWTPs were determined. The results indicated that the distribution characteristics of the different types of contaminants in the three RWTPs were similar. Caffeine, sul2 and tris(1-chloro-2-propyl) phosphate were the dominant substances in the wastewater, and their highest concentrations were 27104 ng/L, 1.4 × 10⁷ copies/mL and 262 ng/L, respectively. Ofloxacin and sul2 were observed to be the dominant substances in the sludge, and their highest concentrations were 5419 ng/g and 3.7 × 10⁸ copies/g, respectively. Anaerobic/anoxic/oxic system combined with the membrane bioreactor process achieved a relatively high aqueous removal of PPCPs (87%). ARGs and OPFRs were challenging to remove, with average removal rates of 6.5% and 31%, respectively. Quantitative meta-analysis indicated that tertiary treatment processes performed better in emerging contaminant removal than secondary processes. Diethyltoluamide exhibited the highest mass load discharge, with 33.5 mg/d per 1000 inhabitants. Octocrylene and tris(2-ethylhexyl) phosphate posed high risks (risk quotient>1.0) to aquatic organisms. This study provides essential evidence to screen high priority pollutants and develop corresponding standard in RWTPs.     

A review on anammox process for the treatment of antibiotic-containing wastewater: Linking effects with corresponding mechanisms

• Anammox is promising for nitrogen removal from antibiotic-containing wastewater. • Most antibiotics could inhibit the anammox performance and activity. • Antibiotic pressure promoted the increase in antibiotic resistance genes (ARGs). • Antibiotic-resistance mechanisms of anammox bacteria are speculated. Antibiotic is widely present in the effluent from livestock husbandry and the pharmaceutical industry. Antibiotics in wastewater usually have high biological toxicity and even promote the occurrence and transmission of antibiotic resistant bacteria and antibiotic resistance genes. Moreover, most antibiotic-containing wastewater contains high concentration of ammonia nitrogen. Improper treatment will lead to high risk to the surrounding environment and even human health. The anaerobic ammonium oxidation (anammox) with great economic benefit and good treatment effect is a promising process to remove nitrogen from antibiotic-containing wastewater. However, antibiotic inhibition has been observed in anammox applications. Therefore, a comprehensive overview of the single and combined effects of various antibiotics on the anammox system is conducted in this review with a focus on nitrogen removal performance, sludge properties, microbial community, antibiotic resistance genes and anammox-involved functional genes. Additionally, the influencing mechanism of antibiotics on anammox consortia is summarized. Remaining problems and future research needs are also proposed based on the presented summary. This review provides a better understanding of the influences of antibiotics on anammox and offers a direction to remove nitrogen from antibiotic-containing wastewater by the anammox process.     

Removal of trimethoprim and sulfamethoxazole in artificial composite soil treatment systems and diversity of microbial communities

• Novel ACST allowed biodegradation to effectively remove adsorbed SMX and TMP. • Ammonia and nitrite were efficiently removed in ACSTs and water quality was improved. Four artificial composite soil treatment systems (ACSTs) fed with reclaimed water containing trimethoprim (TMP) and sulfamethoxazole (SMX) were constructed to investigate SMX and TMP biodegradation efficiency, ammonia and nitrite removal conditions and the microbial community within ACST layers. Results showed SMX and TMP removal rates could reach 80% and 95%, respectively, and removal rates of ammonia and nitrite could reach 80% and 90%, respectively, in ACSTs. The MiSeq sequencing results showed that microbial community structures of the ACSTs were similar. The dominant microbial community in the adsorption and biodegradation layers of the ACSTs contained Proteobacteria, Chloroflexi, Acidobacteria, Firmicutes, Actinobacteria and Nitrospirae. Firmicutes and Proteobacteria were considerably dominant in the ACST biodegradation layers. The entire experimental results indicated that Nitrosomonadaceae_uncultured, Nitrospira and Bacillus were associated with nitrification processes, while Bacillus and Lactococcus were associated with SMX and TMP removal processes. The findings suggest that ACSTs are appropriate for engineering applications.     

Comparison of different valent iron on anaerobic sludge digestion: Focusing on oxidation reduction potential,dissolved organic nitrogen and microbial community

• ORP value from −278.71 to −379.80 mV showed indiscernible effects on methane yield. • Fe(II) and Fe(III) promoted more degradation of proteins and amino acids than Fe⁰. • The highest enrichment of Geobacter was noted in samples added with Fe⁰. • Cysteine was accumulated during iron enhanced anaerobic sludge digestion. • Both iron content and valence were important for methane production. This study compared effects of three different valent iron (Fe⁰, Fe(II) and Fe(III)) on enhanced anaerobic sludge digestion, focusing on the changes of oxidation reduction potential (ORP), dissolved organic nitrogen (DON), and microbial community. Under the same iron dose in range of 0−160 mg/L after an incubation period of 30 days (d), the maximum methane production rate of sludge samples dosed with respective Fe⁰, Fe(II) and Fe(III) at the same concentration showed indiscernible differences at each iron dose, regardless of the different iron valence. Moreover, their behavior in changes of ORP, DON and microbial community was different: (1) the addition of Fe⁰ made the ORP of sludge more negative, and the addition of Fe(II) and Fe(III) made the ORP of sludge less negative. However, whether being more or less negative, the changes of ORP may show unobservable effects on methane yield when it ranged from −278.71 to −379.80 mV; (2) the degradation of dissolved organic nitrogen, particularly proteins, was less efficient in sludge samples dosed with Fe⁰ compared with those dosed with Fe(II) and Fe(III) after an incubation period of 30 d. At the same dose of 160 mg/L iron, more cysteine was noted in sludge samples dosed with Fe(II) (30.74 mg/L) and Fe(III) (27.92 mg/L) compared with that dosed with Fe⁰ (21.75 mg/L); (3) Fe⁰ particularly promoted the enrichment of Geobacter, and it was 6 times higher than those in sludge samples dosed with Fe(II) and Fe(III) at the same dose of 160 mg/L iron.     

Reutilize tire in microbial fuel cell for enhancing the nitrogen removal of the anammox process coupled with iron-carbon micro-electrolysis

• MFC promoted the nitrogen removal of anammox with Fe-C micro-electrolysis. • Reutilize pyrolysis waste tire as micro-electrolysis and electrode materials. • Total nitrogen removal efficiency of modified MFC increased to 85.00%. • Candidatus kuenenia and SM1A02 were major genera responsible for nitrogen removal. In this study, microbial fuel cells (MFCs) were explored to promote the nitrogen removal performance of combined anaerobic ammonium oxidation (anammox) and Fe-C micro-electrolysis (CAE) systems. The average total nitrogen (TN) removal efficiency of the modified MFC system was 85.00%, while that of the anammox system was 62.16%. Additionally, the effective operation time of this system increased from six (CAE system alone) to over 50 days, significantly promoting TN removal. The enhanced performance could be attributed to the electron transferred from the anode to the cathode, which aided in reducing nitrate/nitrite in denitrification. The H⁺ released through the proton exchange membrane caused a decrease in the pH, facilitating Fe corrosion. The pyrolyzed waste tire used as the cathode could immobilize microorganisms, enhance electron transport, and produce a natural Fe-C micro-electrolysis system. According to the microbial community analysis, Candidatus kuenenia was the major genus involved in the anammox process. Furthermore, the SM1A02 genus exhibited the highest abundance and was enriched the fastest, and could be a novel potential strain that aids the anammox process.     

Taxonomic and functional variations in the microbial community during the upgrade process of a full-scale landfill leachate treatment plant – from conventional to partial nitrification-denitrification

• Upgrade process was investigated in a full-scale landfill leachate treatment plant. • The optimization of DO can technically achieve the shift from CND to PND process. • Nitrosomonas was mainly responsible for ammonium oxidation in PND system. • An obviously enrichment of Thauera was found in the PND process. • Enhanced metabolic potentials on organics was found during the process update. Because of the low access to biodegradable organic substances used for denitrification, the partial nitrification-denitrification process has been considered as a low-cost, sustainable alternative for landfill leachate treatment. In this study, the process upgrade from conventional to partial nitrification-denitrification was comprehensively investigated in a full-scale landfill leachate treatment plant (LLTP). The partial nitrification-denitrification system was successfully achieved through the optimizing dissolved oxygen and the external carbon source, with effluent nitrogen concentrations lower than 150 mg/L. Moreover, the upgrading process facilitated the enrichment of Nitrosomonas (abundance increased from 0.4% to 3.3%), which was also evidenced by increased abundance of amoA/B/C genes carried by Nitrosomonas. Although Nitrospira (accounting for 0.1%–0.6%) was found to stably exist in the reactor tank, considerable nitrite accumulation occurred in the reactor (reaching 98.8 mg/L), indicating high-efficiency of the partial nitrification process. Moreover, the abundance of Thauera, the dominant denitrifying bacteria responsible for nitrite reduction, gradually increased from 0.60% to 5.52% during the upgrade process. This process caused great changes in the microbial community, inducing continuous succession of heterotrophic bacteria accompanied by enhanced metabolic potentials toward organic substances. The results obtained in this study advanced our understanding of the operation of a partial nitrification-denitrification system and provided a technical case for the upgrade of currently existing full-scale LLTPs.     

Microbial responses to the use of NaClO in sediment treatment

• Chlorine addition enhanced the release of TOC, TN from the sediment. • Chlorine has a long-term negative effect on microbial richness. • Usually enzymes lose activity, and expression of genes was downregulated. • Carbon degradation and nitrification might be strongly inhibited. Chlorine is often used in algal removal and deodorization of landscape waters, and occasionally used as an emergency treatment of heavily polluted sediments. However, the ecological impact of this practice has not been fully studied and recognized. In this study, NaClO at 0.1 mmol/g based on dry weight sediment was evenly mixed into the polluted sediment, and then the sediment was incubated for 150 days to evaluate its microbial effect. Results showed that NaClO addition enhanced the release of TOC, TN, Cr and Cu from the sediment. The microbial richness in the examined sediment decreased continuously, and the Chao1 index declined from 4241 to 2731, in 150 days. The microbial community composition was also changed. The abundance of Proteobacteria and Bacteroidetes increased to 54.8% and 4.2% within 7 days compared to the control, and linear discriminant analysis (LDA) showed gram-negative bacteria and aerobic bacteria enriched after chlorination. The functional prediction with PICRUSt2 showed the functions of the microbial community underwent major adjustments, and the metabolic-related functions such as carbon metabolism, including pyruvate and methane metabolisms were significantly inhibited; besides, 15 out of 22 analyzed key enzymes involved in C cycling and 6 out of 12 key enzymes or genes involved in N cycling were strongly impacted, and the enzymes and genes involved in carbon degradation and denitrification showed remarkable downregulation. It can be concluded that chlorination posed a seriously adverse effect on microbial community structure and function. This study deepens the understanding of the ecological effects of applying chlorine for environmental remediation.     

Mechanisms for simultaneous ozonation of sulfamethoxazole and natural organic matters in secondary effluent from sewage treatment plant

• SMX was mainly degraded by hydrolysis, isoxazole oxidation and double-bond addition. • Isoxazole oxidation and bond addition products were formed by direct ozonation. • Hydroxylated products were produced by indirect oxidation. • NOM mainly affected the degradation of SMX by consuming ^•OH rather than O₃. • Inhibitory effect of NOM on SMX removal was related to the components’ aromaticity. Sulfamethoxazole (SMX) is commonly detected in wastewater and cannot be completely decomposed during conventional treatment processes. Ozone (O₃) is often used in water treatment. This study explored the influence of natural organic matters (NOM) in secondary effluent of a sewage treatment plant on the ozonation pathways of SMX. The changes in NOM components during ozonation were also analyzed. SMX was primarily degraded by hydrolysis, isoxazole-ring opening, and double-bond addition, whereas hydroxylation was not the principal route given the low maximum abundances of the hydroxylated products, with m/z of 269 and 287. The hydroxylation process occurred mainly through indirect oxidation because the maximum abundances of the products reduced by about 70% after the radical quencher was added, whereas isoxazole-ring opening and double-bond addition processes mainly depended on direct oxidation, which was unaffected by the quencher. NOM mainly affected the degradation of micropollutants by consuming ^•OH rather than O₃ molecules, resulting in the 63%–85% decrease in indirect oxidation products. The NOM in the effluent were also degraded simultaneously during ozonation, and the components with larger aromaticity were more likely degraded through direct oxidation. The dependences of the three main components of NOM in the effluent on indirect oxidation followed the sequence: humic-like substances>fluvic-like substances>protein-like substances. This study reveals the ozonation mechanism of SMX in secondary effluent and provides a theoretical basis for the control of SMX and its degradation products in actual water treatment.     

PICRUSt2 functionally predicts organic compounds degradation and sulfate reduction pathways in an acidogenic bioreactor

• Over 70% reduction of sulfate was achieved for sulfate less than 12000 mg/L. • The decrease of genes encoding (EC: 1.3.8.1) induced the accumulation of VFAs. • The sulfate reduction genes were primary carried by genus Desulfovibrio. • Sulfate favored assimilatory, but inhibited dissimilatory sulfate reduction process. For comprehensive insights into the influences of sulfate on performance, microbial community and metabolic pathways in the acidification phase of a two-phase anaerobic system, a laboratory-scale acidogenic bioreactor was continuously operated to treat wastewater with elevated sulfate concentrations from 2000 to 14000 mg/L. The results showed that the acidogenic bioreactor could achieve sulfate reduction efficiency of greater than 70% for influent sulfate content less than 12000 mg/L. Increased sulfate induced the accumulation of volatile fatty acids (VFAs), especially propionate and butyrate, which was the primary negative effects to system performance under the high-sulfate environment. High-throughput sequencing coupled with PICRUSt2 uncovered that the accumulation of VFAs was triggered by the decreasing of genes encoding short-chain acyl-CoA dehydrogenase (EC: 1.3.8.1), regulating the transformation of propanoyl-CoA to propenoyl-CoA and butanoyl-CoA to crotonyl-CoA of propionate and butyrate oxidation pathways, which made these two process hardly proceed. Besides, genes encoding (EC: 1.3.8.1) were mainly carried by order Clostridiales. Desulfovibrio was the most abundant sulfate-reducing bacteria and identified as the primary host of dissimilatory sulfate reduction functional genes. Functional analysis indicated the dissimilatory sulfate reduction process predominated under a low sulfate environment, but was not favored under the circumstance of high-sulfate. With the increase of sulfate, the assimilatory sulfate reduction process finally overwhelmed dissimilatory as the dominant sulfate reduction pathway in acidogenic bioreactor.