Microplastics Enhance the Prevalence of Antibiotic Resistance Genes in Anaerobic Sludge Digestion by Enriching Antibiotic-Resistant Bacteria in Surface Biofilm and Facilitating the Vertical and Horizontal Gene Transfer.

Antibiotic resistance genes (ARGs) and microplastics (MPs) are recognized as emerging contaminants and threats to global human health. Despite both of them being significantly detected in their "hotspots", i.e., waste activated sludge (WAS), rare studies on how MPs affect ARGs and antibiotic-resistant bacteria (ARB) in anaerobic sludge digestion are available. Herein, the fate of ARGs and ARB after exposure to MPs of three dosages (10, 30, and 80 particles/g-TS), three polymer types (LDPE, PET, and PS), and three branching extents (LDPE, LLDPE, and HDPE) in anaerobic sludge digestion was investigated. Metagenomic results indicated that all variants of MPs resulted in an increase of the relative abundance of ARGs in the digester compared to the control. The abundance of ARGs demonstrated a dosage-dependent relationship within the range from 10 to 80 particles/g-TS, resulting in an increase from 4.5 to 27.9% compared to the control. Branching structure and polymer type influence ARG level in the sludge digester as well. Mechanism studies revealed that LDPE selectively enriched potential ARB and ARGs in the surface biofilm, possibly creating a favorable environment for ARB proliferation and ARG exchange. Furthermore, vertical transfer of ARGs was facilitated by LDPE through increasing bacterial cell proliferation accompanied by the enhancement of relevant functional genes. The elevated abundance of mobile genetic elements (MGEs) and ARGs-carrying plasmids also demonstrated that MGE-mediated horizontal transfer was promoted by LDPE at 80 particles/g-TS. This effect was compounded by increased oxidative stress, cell membrane permeability, and cell cohesion, collectively facilitating horizontal ARG transfer. Consequently, both vertical and horizontal transfer of ARGs could be concurrently promoted by LDPE an in anaerobic sludge digester.

Environmentally relevant level of perfluorooctanoic acid affect the formation of aerobic granular sludge.

The growing increasing occurrence of perfluorooctanoic acid (PFOA) in wastewater has raised concerns about its potential impact on the environment. Nevertheless, the impact of PFOA at environmentally relevant level on the formation of aerobic granular sludge (AGS) is still a 'black box'. This study thus aims to fill this gap by comprehensive investigation of sludge properties, reactor performance and microbial community during the formation of AGS. It was found that 0.1 mg/L PFOA delayed the formation of AGS, causing relatively lower proportion of large size AGS at the end of operation process. Interestingly, the microorganisms contribute to the reactor's tolerance to PFOA by secreting more extracellular polymeric substances (EPS) to slow or block the entry of toxic substances into the cells. During the granule maturation period, the reactor nutrient removal especially chemical oxygen demand (COD) and total nitrogen (TN) were affected by PFOA, decreasing the corresponding removal efficiencies to ∼81.2% and ∼69.8%, respectively. Microbial analysis further revealed that PFOA decreased the abundances of Plasticicumulans, Thauera, Flavobacterium and Cytophagaceae_uncultured, but it has promoted Zoogloea and Betaproteobacteria_unclassified growth, which maintained the structures and functions of AGS. The above results revealed that the intrinsic mechanism of PFOA on the macroscopic representation of sludge granulation process was revealed, and it is expected to provide theoretical insights and practical support for direct adoption of municipal or industrial wastewater containing perfluorinated compounds to cultivate AGS.

Effects of pneumatic compression therapy on wound healing in patients with venous ulcers: A meta-analysis.

This meta-analysis assessed the effect of pneumatic compression therapy on the wound healing of venous ulcers, with the aim of providing a basis for the selection of clinical treatment. Randomised controlled trials (RCTs) on the application of pneumatic compression therapy to venous ulcers were collected by searching PubMed, Embase, Cochrane Library, China National Knowledge Infrastructure, VIP, and Wanfang databases, with a timeframe from database inception to August 2023. After two researchers independently screened the literature, extracted information, and evaluated the quality of the included studies, a meta-analysis was performed using RevMan 5.4 software. Six RCTs with 367 patients were included, with 172 patients in the intervention group and 195 in the control group. The results showed that pneumatic and bandage compression therapies had a similar impact on wound healing rates of venous ulcers (54.65% vs. 53.84%, odds ratio [OR]: 1.02, 95% confidence interval [CI]: 0.49-2.12, p = 0.96), changes in wound area (standardised mean difference: -0.16, 95% CIs: -0.45 to 0.12, p = 0.26), adverse event rates (76.56% vs. 67.07%, OR: 1.62, 95% CI: 0.77-3.39, p = 0.20), and the differences were not statistically significant. Thus, current evidence suggests that the effects of pneumatic compression therapy on wound healing rates, changes in wound area, and the incidence of adverse events in patients with venous ulcers are similar to those of bandage pressure therapy. However, owing to the limitations in the number and quality of studies, more high-quality RCTs are needed to clarify the feasibility and economics of pneumatic compression therapy in patients with venous ulcers.

Performance and Mechanism of Fe3O4 Improving Biotransformation of Waste Activated Sludge into Liquid High-Value Products.

This study demonstrated that Fe3O4 simultaneously improves the total production and formation rate of medium-chain fatty acids (MCFAs) and long-chain alcohols (LCAs) from waste activated sludge (WAS) in anaerobic fermentation. Results revealed that when Fe3O4 increased from 0 to 5 g/L, the maximal MCFA and LCA production increased significantly, and the optimal fermentation time was also remarkably shortened from 24 to 9 days. Moreover, Fe3O4 also enhanced WAS degradation, and the corresponding degradation rate in the fermentation system increased from 43.86 to 72.38% with an increase in Fe3O4 from 0 to 5 g/L. Further analysis showed that Fe3O4 promoted the microbe activities of all the bioprocesses (including hydrolysis, acidogenesis, and chain elongation processes) involved in the MCFA and LCA production from WAS. Microbial community analysis indicated that Fe3O4 increased the abundances of key microbes involved in abovementioned bioprocesses correspondingly. Mechanistic investigations showed that Fe3O4 increased the conductivity of the fermented sludge, providing a better conductive environment for the anaerobic microbes. The redox cycle of Fe(II) and Fe(III) existed in the fermentation system with Fe3O4, which was likely to act as electron shuttles to conduct electron transfer (ET) from the electron donor to the acceptor, thus increasing ET efficiency. This study provides an effective method for enhancing the biotransformation of WAS into high-value products, potentially bringing economic benefits to WAS treatment.

α-Pinene inhibits the growth of cervical cancer cells through its proapoptotic activity by regulating the miR-34a-5p/Bcl-2 signaling axis.

Among gynecological tumors, cervical cancer (CC) has the second-highest prevalence and mortality rate. α-Pinene is a bicyclic monoterpenoid compound extracted from pine needles that carried promising anticancer properties. Nevertheless, its effect on CC and the underlying mechanism has not yet been elucidated. Therefore, we investigated the effect of α-Pinene on apoptosis in CC via in vitro assays of flow cytometry (FCW), terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) assay, quantitative real-time polymerase chain reaction (qRT-PCR), and Western blot. Following that, we detected the proapoptotic function of α-Pinene on HeLa cells in vivo by TUNEL assay and immunofluorescence staining. Our results displayed that the α-Pinene inhibited the growth of HeLa cells and stalled the cells in the G0/G1 phase. Interestingly, we also detected that α-Pinene induced HeLa cells to apoptosis. The results investigated that α-Pinene induced HeLa cells apoptosis along with up-regulating the expression of Bax, Bid, caspase-9, caspase-3, miR-34a-5p, and down-regulating the expression of Bcl-2 in vitro. At the same time, the expression levels of target genes in vivo were consistent with those in vitro. Our experiment proved that α-Pinene promoted apoptosis, which will be used to hopefully maximize the therapeutic strategies in clinical studies in CC.

Biological Reduction of Nitric Oxide for Efficient Recovery of Nitrous Oxide as an Energy Source.

Chemical absorption-biological reduction based on Fe(II)EDTA is a promising technology to remove nitric oxide (NO) from flue gases. However, limited effort has been made to enable direct energy recovery from NO through production of nitrous oxide (N2O) as a potential renewable energy rather than greenhouse gas. In this work, the enhanced energy recovery in the form of N2O via biological NO reduction was investigated by conducting short-term and long-term experiments at different Fe(II)EDTA-NO and organic carbon levels. The results showed both NO reductase and N2O reductase were inhibited at Fe(II)EDTA-NO concentration up to 20 mM, with the latter being inhibited more significantly, thus facilitating N2O accumulation. Furthermore, N2O accumulation was enhanced under carbon-limiting conditions because of electron competition during short-term experiments. Up to 47.5% of NO-N could be converted to gaseous N2O-N, representing efficient N2O recovery. Fe(II)EDTA-NO reduced microbial diversity and altered the community structure toward Fe(II)EDTA-NO-reducing bacteria-dominated culture during long-term experiments. The most abundant bacterial genus Pseudomonas, which was able to resist the toxicity of Fe(II)EDTA-NO, was significantly enriched, with its relative abundance increased from 1.0 to 70.3%, suggesting Pseudomonas could be the typical microbe for the energy recovery technology in NO-based denitrification.

The impact and fate of clarithromycin in anaerobic digestion of waste activated sludge for biogas production.

Clarithromycin retained in waste activated sludge (WAS) inevitably enters the anaerobic digestion system. So far, the complex impacts and fate of clarithromycin in continuous operated WAS anaerobic digestion system are still unclear. In this study, two semi-continuous long-term reactors were set up to investigate the effect of clarithromycin on biogas production and antibiotic resistance genes (ARGs) during WAS anaerobic digestion, and a batch test was carried out to explore the potential metabolic mechanism. Experimental results showed that clarithromycin at lower concentrations (i.e., 0.1 and 1.0 mg/L) did not affect biogas production, whereas the decrease in biogas production was observed when the concentration of clarithromycin was further increased to 10 mg/L. Correspondingly, the relative abundance of functional bacteria in WAS anaerobic digestion (i.e., Anaerolineaceae and Microtrichales) was reduced with long-term clarithromycin exposure. The investigation of ARGs suggested that the effect of methylation belonging to the target site modification played a critical role for the anaerobic microorganisms in the expression of antibiotic resistance, and ermF, played dominated ARGs, presented the most remarkable proliferation. In comparison, the role of efflux pump was weakened with a significant decrease of two detected efflux genes. During WAS anaerobic digestion, clarithromycin could be partially degraded into metabolites with lower antimicrobial activity including oleandomycin and 5-O-desosaminyl-6-O-methylerythronolide and other metabolites without antimicrobial activity.

Long-Term Effects of Polyvinyl Chloride Microplastics on Anaerobic Granular Sludge for Recovering Methane from Wastewater.

Polyvinyl chloride microplastics (PVC-MPs) are emerging contaminants affecting biological wastewater treatment processes. However, most of the previous studies mainly focused on their short-term impacts on floc sludge, with little work being conducted to explore their potential effects on more complex anaerobic granular sludge (AGS), which has been widely used for high-strength organic wastewater treatment. In this paper, the long-term effects of PVC-MPs on AGS were investigated via continuous feeding tests that are representative of real wastewater treatment processes. The results of a continuous 264 days test showed that the prolonged exposure of PVC-MPs at 15-150 MPs·L-1 significantly (p = 7.86 × 10-37, 3.44 × 10-43, and 5.29 × 10-46) inhibited the COD removal efficiency of AGS by 13.2%-35.5%, accompanied by a 11.0%-32.3% decreased production of methane and 40.3%-272.7% increased accumulation of short-chain fatty acids (SCFAs). In addition, the PVC-MPs exposure suppressed the secretion of extracellular polymeric substances (EPS), causing AGS and the inside microorganisms to lose the protection of EPS, thereby resulting in granule breakage and decreased cells viability. Aligning with the deteriorated performance, the long-term exposure of PVC-MPs reduced the total microbial populations and the relative abundances of key methanogens and acidogens. A toxicity mechanism assessment revealed that the negative impacts induced by PVC-MPs are mainly attributed to the toxic leachate and excess oxidative stress.

Hydrated lanthanum oxide-modified diatomite as highly efficient adsorbent for low-concentration phosphate removal from secondary effluents.

The requirement to the phosphorus (P) emission from wastewater treatment plants (WWTPs) is becoming increasingly strict, which makes an advanced treatment for the low-concentration phosphate removal from secondary effluents indispensable. In present work, hydrated lanthanum (La) oxide-modified diatomite composites (La-diatomite) were fabricated by a facile method and employed as the highly efficient adsorbent for the low-concentration phosphate removal from simulating secondary effluents. Comparative experiments indicated that the La-diatomite treated by 0.1 mol/L LaCl3 exhibited the highest La availability (P/La molar ratio of 2.30) and performed good selectivity to phosphate adsorption even with the coexistence of competing anions and humic acid. The maximum P adsorption capacity reached to 58.7 mg P/g and the 96% P was removed quickly within 30 min at initial phosphate concentration 2 mg P/L. Insignificant La leaching was observed during the process due to the La stabilization by macroporous diatomite. Eight cycles of adsorption-desorption experiments revealed that the excellent repeated use property of La-diatomite. At the column test, La-diatomite showed superior treatment capacities of 3455 kg water/kg La-diatomite for simulated secondary effluents. The La-diatomite maintained high and stable adsorption effectiveness in wide pH range, which should be attributed to the synergistic effect of electrostatic interactions, ligand exchange and Lewis acid-based interaction. This work might provide a candidate for low-concentration phosphate removal from secondary effluent to alleviate the eutrophication.

The fate of cyanuric acid in biological wastewater treatment system and its impact on biological nutrient removal.

Cyanuric acid (CA) is widely used in living and production. It is a kind of environmental priority pollutants which exists chronically in soil and water, but is difficult to be chemically hydrolyzed or oxidized. The behavior of CA at different levels of 0, 0.01, 0.10 and 1.00 mg L-1 in biological wastewater treatment process was investigated in this paper. Experimental results showed that CA (0.01 and 0.10 mg L-1) was removed in biological wastewater treatment process, which was mainly achieved by biodegradation of particular species (Acidovorax and Pseudomonas) in the anaerobic condition. However, 1.00 mg L-1 CA was reluctant to be degraded in biological wastewater treatment system. With the CA level increase from 0 to 1.00 mg L-1, total nitrogen removal efficiency decreased from 97.23 to 74.72%. The presence of CA promoted both the synthesis and decomposition metabolisms of poly-hydroxyalkanoates and glycogen, thereby providing the advantage for phosphorus removal. CA could inhibit nitrification process because of inhibition to nitrite oxidizing bacteria (NOB). Moreover, the microbial community of activated sludge was changed by the exposure of CA. Polyphosphate accumulating organisms, such as Bacteroidetes, Chloroflexi and Saccharibacteria increased, but the abundance of Nitrospirae was decreased.

[Synthesis, biological activity, computer aided drug design of alpha-pinene derivatives].

Based on the anticancer mechanism of biological alkylating agent, we designed and synthesized two alpha pinene derivatives:(1R,5S)-(6,6-dimethylbicyclo[3,1,1]hept-2-en-2-yl)methyl benzenesulfonate and (1R,5S)-(6,6-dimethylbicyclo[3,1,1]hept-2-en-2-yl)methyl 4-methylbenzenesulfonate, of which structures were confirmed by ¹H-NMR, HPLC and MS date. These two compounds showed a good inhibition of tumor cells' proliferation. Further, the computer siuulation of molecular docking and metabolic kinetics indicated that these two copounds may have stable molecular complexation with protein CDK2, which closely related to the cell cycle.

Effects of mechanical stretching on the morphology and cytoskeleton of vaginal fibroblasts from women with pelvic organ prolapse.

Mechanical load and postmenopausal hypoestrogen are risk factors for pelvic organ prolapse (POP). In this study, we applied a 0.1-Hz uniaxial cyclic mechanical stretching (CS) with 10% elongation and 10⁻8 M 17-ß-estradiol to vaginal fibroblasts isolated from postmenopausal women with or without POP to investigate the effects of CS and estrogen on cell morphology and cytoskeletons of normal and POP fibroblasts. Under static culture condition, POP fibroblasts exhibited lower cell circularity and higher relative fluorescence intensities (RFIs) of F-actin, α-tubulin and vimentin. When cultured with CS, all fibroblasts grew perpendicular to the force and exhibited a decreased cell projection area, cell circularity and increased cell length/width ratio; normal fibroblasts exhibited increased RFIs of all three types of cytoskeleton, and POP fibroblasts exhibited a decreased RFI of F-actin and no significant differences of α-tubulin and vimentin. After being cultured with 17-ß-estradiol and CS, normal fibroblasts no longer exhibited significant changes in the cell projection area and the RFIs of F-actin and α-tubulin; POP fibroblasts exhibited no significant changes in cell circularity, length/width ratio and F-actin even with the increased RFIs of α-tubulin and vimentin. These findings suggest that POP fibroblasts have greater sensitivity to and lower tolerance for mechanical stretching, and estrogen can improve the prognosis.

[Comparative study on toxicity of extracts from Phytolaccae Radix before and after being processed with vinegar].

OBJECTIVE: To extract and separate toxic components from Phytolaccae Radix, and to comare the changes in toxicity of Phytolaccae Radix before and after being processed with vinegar. METHOD: The mucous membrane irritation response, mouse peritoneal inflammation model and in vitro macrophages release NO model were applied to compared the changes in inflammatory toxicity of toxic components from Phytolaccae Radix before and after being processed with vinegar. RESULT: Toxic components of Phytolacca Radix had significant inflammatory toxicity, which could cause conjunctival edema in rabbits, and increase of PGE2 and macrophages release NO content in peritoneal exudate in mice. After being processed with vinegar, they showed reduced irritation, which resulted in decrease of PGE2 and macrophages release NO content in peritoneal exudate in mice. CONCLUSION: After being processed with vinegar, the toxicity of toxic components from Phytolacca Radix decreased obviously.

Hydrochar mediated anaerobic digestion of bio-wastes: Advances, mechanisms and perspectives.

Bio-wastes treatment and disposal has become a challenge because of their increasing output. Given the abundant organic matter in bio-wastes, its related resource treatment methods have received more and more attention. As a promising strategy, anaerobic digestion (AD) has been widely used in the treatment of bio-wastes, during which not only methane as energy can be recovered but also their reduction can be achieved. However, AD process is generally disturbed by some internal factors (e.g., low hydrolysis efficiency and accumulated ammonia) and external factors (e.g., input pollutants), resulting in unstable AD operation performance. Recently, hydrochar was wildly found to improve AD performance when added to AD systems. This review comprehensively summarizes the research progress on the performance of hydrochar-mediated AD, such as increased methane yield, improved operation efficiency and digestate dewatering, and reduced heavy metals in digestate. Subsequently, the underlying mechanisms of hydrochar promoting AD were systematically elucidated and discussed, including regulation of electron transfer (ET) mode, microbial community structure, bio-processes involved in AD, and reaction conditions. Moreover, the effects of properties of hydrochar (e.g., feedstock, hydrothermal carbonization (HTC) temperature, HTC time, modification and dosage) on the improvement of AD performance are systematically concluded. Finally, the relevant knowledge gaps and opportunities to be studied are presented to improve the progress and application of the hydrochar-mediated AD technology. This review aims to offer some references and directions for the hydrochar-mediated AD technology in improving bio-wastes resource recovery.

Different microplastics distinctively enriched the antibiotic resistance genes in anaerobic sludge digestion through shifting specific hosts and promoting horizontal gene flow.

Both microplastics (MPs) and antibiotic resistance genes (ARGs) are intensively detected in waste activated sludge (WAS). However, the distinctive impacts of different MPs on ARGs emergence, dissemination, and its potential mechanisms remain unclear. In this study, long-term semi-continuous digesters were performed to examine the profiles of ARGs and antibiotic-resistant bacteria (ARB) in response to two different typical MPs (polyethylene (PE) and polyvinyl chloride (PVC)) in anaerobic sludge digestion. Metagenomic results show that PE- and PVC-MPs increase ARGs abundance by 14.8% and 23.6% in digester, respectively. ARB are also enriched by PE- and PVC-MPs, Acinetobacter sp. and Salmonella sp. are the dominant ARB. Further exploration reveals that PVC-MPs stimulates the acquisition of ARGs by human pathogen bacteria (HPB) and functional microorganisms (FMs), but PE-MPs doesn't. Network analysis shows that more ARGs tend to co-occur with HBP and FMs after MPs exposure, and more importantly, new bacteria are observed to acquire ARGs possibly via horizontal gene flow (HGF) in MPs-stressed digester. The genes involved in the HGF process, including reactive oxygen species (ROS) production, cell membrane permeability, extracellular polymeric substances (EPS) secretion, and ATP synthesis, are also enhanced by MPs, thereby attributing to the promoted ARGs dissemination. These findings offer advanced insights into the distinctive contribution of MPs to fate, host, dissemination of ARGs in anaerobic sludge digestion.

[Comparative study on toxicity of Euphorbia before and after being prepared by vinegar].

OBJECTIVE: To study and compare the changes of toxicity of Euphorbia pekinensis, E. kansui and E. ebracteolata before and after being prepared by vinegar. METHOD: Small intestinal accentuation of mice and peritoneal macrophage NO release experiments were assessed to investigate the changes of toxicity of the three Chinese Medicines of Euphorbia before and after being prepared. RESULT: E. pekinensis, E. kansui and E. ebracteolata and vinegar can obviously promot small intestinal accentuation and peritoneal macrophage NO release with the intensity of toxicity in the order of E. kansui > E. pekinensis > E. ebracteolata. After being prepared with vinegar, the toxicity of the three medicines decreased obviously compared to crude one. CONCLUSION: E. pekinensis, E. kansui and E. ebracteolata can induce inflammation and accelerate enterokinesis. After being prepared with vinegar, the irritation on Euphorbia decreased obviously.

Revealing the mechanisms of rhamnolipid enhanced hydrogen production from dark fermentation of waste activated sludge.

Rhamnolipid (RL), as an environmentally compatible biosurfactant, has been used to enhance waste activated sludge (WAS) fermentation. However, the effect of RL on hydrogen accumulation in anaerobic fermentation remains unclear. Therefore, this work targets to investigate the mechanism of RL-based dark fermentation system on hydrogen production of WAS. It was found that the maximum yield of hydrogen increased from 1.76 ± 0.26 to 11.01 ± 0.30 mL/g VSS (volatile suspended solids), when RL concentration increased from 0 to 0.10 g/g TSS (total suspended solids). Further enhancement of RL level to 0.12 g/g TSS slightly reduced the production to 10.80 ± 0.28 mL/g VSS. Experimental findings revealed that although RL could be degraded to generate hydrogen, it did not play a major role in enhancing hydrogen accumulation. Mechanism analysis suggested that RL decreased the surface tension between sludge liquid and hydrophobic compounds, thus accelerating the solubilization of WAS, improving the proportion of biodegradable substances which could be used for subsequent hydrogen production. Regardless of the fact that adding RL suppressed all the fermentation processes, the inhibition effect of processes associated with hydrogen consumption was much severer than that of hydrogen production. Further investigations of microbial community revealed that RL enriched the relative abundance of hydrogen producers e.g., Romboutsia but reduced that of hydrogen consumers like Desulfobulbus and Caldisericum.

Hydrochar prepared from digestate improves anaerobic co-digestion of food waste and sewage sludge: Performance, mechanisms, and implication.

This work reported a new waste functionalization and utilization method, which use digestate to prepare hydrochar to improve methane production from food waste (FW) and sewage sludge (SS). Experimental results presented that 10 g/L hydrochar obtained the cumulative methane production of 133.11 ± 1.18 mL/g volatile solids added, 26.99 % higher than that without hydrochar addition. By monitoring the conversion of model metabolic intermediates, 10 g/L hydrochar was determined to favor hydrolysis, acidogenesis and methonogenesis bio-processes involved in methane production, thus improving the degradation of solubilized organics and consumption of short-chain fatty acids (SCFAs) during the co-digestion. Microbial investigation revealed that 10 g/L hydrochar enriched the microbes relevant to methane production (e.g., Methanosaeta and Syntrophomonas), but reduced the abundances of hydrolysis- and acidogenesis-related microbes (e.g., Acinetobacter). This hydrochar-based preparation and utilization strategy might offer a novel paradigm for waste-control-waste, bringing economic and environmental benefits.

Rhamnolipid pretreatment enhances methane production from two-phase anaerobic digestion of waste activated sludge.

In this work, a rhamnolipid (RL) pretreatment technology was proposed to promote methane production from two-phase anaerobic digestion of waste activated sludge. In the first phase (i.e., acidogenic phase), the WAS hydrolysis and acidogenesis were significantly enhanced after RL pretreatment for 4 day, under which the concentration of soluble protein and the short-chain fatty acids (SCFA) in the presence of RL at 0.04 g/g TSS was respectively 2.50 and 5.02 times higher than that without RL pretreatment. However, methane production was inhibited in the presence of RL. In the second phase (i.e., methanogenic phase), batch biochemical methane potential tests suggested that the addition of RL is effective in promoting anaerobic methane production. With an increase of RL dosage from 0 to 0.04 g/g TSS, the cumulative methane yield increased from 100.42 ± 3.01 to 168.90 ± 5.42 mL. Although the added RL could be utilized to produce methane, it was not the major contributor to the enhancement of methane yield. Further analysis revealed that total cumulative yield from the entire two-phase anaerobic digestion (sum of the yield of the acidogenic phase and methanogenic phase) increased from 113.42 ± 3.56 to 164.18 ± 5.20 mL when RL dosage increased from 0 to 0.03 g/g TSS, indicating that the addition of RL induced positive effect on the methane production of the entire two-phase anaerobic digestion. The enzyme activity analysis showed that although higher dosages of RL still inhibited the microorganisms related to methanogenesis to some extends in the methanogenic phase, the inhibitory effect was significantly weakened compared to the acidogenic phase. Microbial analysis revealed that RL reduced the abundance of Candidatus_Methanofastidiosum sp. while increased the abundance of Methanosaeta sp., causing the major methanogenesis pathway to change from hydrogenotrophic to aceticlastic. Moreover, the community of hydrolytic microbes and acidogens was shifted in the direction that is conducive to hydrolysis-acidogenesis. The findings reported not only expand the application field of RL, but also may provide supports for sustainable operation of wastewater treatment plants (WWTPs).

Benzyl butyl phthalate activates prophage, threatening the stable operation of waste activated sludge anaerobic digestion.

The stable operation of the anaerobic digestion of waste activated sludge (WAS) is threatened by numerous emerging contaminants. Meanwhile, the extensive microplastic pollution increased the environmental exposure risk of plasticizer benzyl butyl phthalate (BBP), the BBP content has reached a substantial level in WAS. However, the effect of BBP on WAS anaerobic digestion is still unknown. Here we show that high-level BBP brings on anaerobic digestion upset. The presence of 10.0 mg/L BBP (in sludge with 17,640 ± 510 mg/L TSS) led to deferred cell lysis, which was confirmed by the results of continuous parallel factor analysis of dissolved organic matter and the liberation of lactate dehydrogenase. Further, the deferred cell rupture was confirmed associate with prophage activation during WAS anaerobic digestion. Besides solubilization, the hydrolysis, acetogenesis and methanogenesis were also affected by the addition of BBP. The long-term effects of BBP revealed that the dominant microbial structure in anaerobic digester was stable, but the abundance of many functional microorganisms was changed, including short chain fatty acid producers and consumers. This work highlights one of the susceptibility mechanisms for WAS anaerobic digestion processes and provides new perspectives for the comprehensive assessment of emerging contaminant's environmental risks.