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Pharmaceutical waste sludge harbors large amounts of antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs), and it is necessary to study the reduction of ARGs and MGEs during sludge treatment. Therefore, the antibiotic resistance phenotypes and genotypes of enterococci, and the ARGs and MGEs in genomic DNA were investigated during anaerobic digestion (AD) with microwave (MW), thermal hydrolysis (TH) and ozone pretreatment. Results showed that sludge pretreatment increased the occurrence of the resistance phenotypes and genotypes of enterococci. During AD, the resistance of enterococci to macrolides decreased, except for in the MW-pretreated sludge. Horizontal gene transfer and co-occurrence of erm B and tet M in enterococci resulted in increased tetracycline resistance of enterococci throughout the sludge treatment. MGEs such as intI 1, IS CR1 and Tn 916/1545 had a significant effect on the distribution of ARGs. AD with pretreatment, especially TH pretreatment, resulted in greater ARGs and MGEs reduction and improved methane production. [ABSTRACT FROM AUTHOR]

The fate of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) were investigated during the sludge anaerobic digestion (AD) with microwave-acid (MW-H), microwave (MW) and microwave-H 2 O 2 -alkaline (MW-H 2 O 2 ) pretreatments. Results showed that combined MW pretreatment especially for the MW-H pretreatment could efficiently reduce the ARB concentration, and most ARG concentrations tended to attenuate during the pretreatment. The subsequent AD showed evident removal of the ARB, but most ARGs were enriched after AD. Only the concentration of tet X kept continuous declination during the whole sludge treatment. The total ARGs concentration showed significant correlation with 16S rRNA during the pretreatment and AD. Compared with unpretreated sludge, the AD of MW and MW-H 2 O 2 pretreated sludge presented slightly better ARB and ARGs reduction efficiency. [ABSTRACT FROM AUTHOR]

Swine farm and the adjacent farmland are hot spots of ARGs. However, few studies have investigated the on-site occurrence of ARGs distributed in the process of anaerobic digestion (AD) followed by land application of swine wastewater. Two typical swine farms, in southern and northern China respectively, with AD along with land application were explored on ARG distributions. ARGs were highly abundant in raw swine wastewater, AD effectively reduced the copy number of all detected ARGs (0.21–1.34 logs removal), but the relative abundance with different resistance mechanisms showed distinctive variation trends. The reduction efficiency of ARGs was improved by stable operational temperature and longer solid retention time (SRT) of AD. ARGs in soil characterized the contamination from the irrigation of the digested liquor. The total ARGs quantity in soil fell down by 1.66 logs in idle period of winter compared to application period of summer in the northern region, whereas the total amount was steady with whole-year application in south. Some persistent ( sul 1 and sul 2) and elevated ARGs ( tet G and ere A) in AD and land application need more attention. [ABSTRACT FROM AUTHOR]

Abstract: The effect of addition of organic carbon sources (acetic acid and waste activated sludge alkaline fermentation liquid) on anaerobic–aerobic (low dissolved oxygen, 0.15–0.45mg/L) biological municipal wastewater treatment was investigated. The results showed that carbon source addition affected not only the transformations of polyhydroxyalkanoates (PHA), glycogen, nitrogen and phosphorus, but the net removal of nitrogen and phosphorus. The removal efficiencies of TN and TP were, respectively, 61% and 61% without organic carbon source addition, 81% and 95% with acetic acid addition, and 83% and 97% with waste activated sludge alkaline fermentation liquid addition. It seems that the alkaline fermentation liquid of waste biosolids generated in biological wastewater treatment plant can be used to replace acetic acid as an additional carbon source to improve the anaerobic–aerobic (low dissolved oxygen) municipal wastewater nutrients removal although its use was observed to cause a slight increase of effluent BOD and COD concentrations. [Copyright &y& Elsevier]

• Dominant bacteria shifted from nutrients removal bacteria to fermentative bacteria. • Hydrogenotrophic pathway is the main biochemical pathway of methanogenesis in AD. • Full-scale thermal hydrolysis & AD selected erm B, erm F, mef A/E, qnr S and tet M. • Archaea biomass and community presented important impacts on the fate of ARGs. • Total ARGs in the sludge are mainly affected by MGEs during the advanced AD. Anaerobic digestion (AD) with thermal hydrolysis pretreatment is widely used as an efficient sludge treatment nowadays. However, the evolution of microbial community (especially for the archaea community), the fate of antibiotic resistance genes (ARGs), and their associations during such process in full-scale sludge treatment plants are rarely reported. Therefore, these scientific questions were explored at two full-scale sludge treatment plants through high-throughput sequencing and quantitative PCR. Results showed that Methanobacterium and Methanosphaera were the dominant archaea in thermal hydrolyzed sludge. The predominant bacteria in the sludge first shifted from nutrients removal functional bacteria to spore-forming bacteria after thermal hydrolysis, and then shifted to fermentative bacteria after AD. The full-scale plants could select erm B, erm F, mef A/E, qnr S and tet M. Though the bacteria and archaea biomass and community largely influenced the fate of ARGs, multiple linear regression analysis showed that the total ARGs were mainly affected by mobile genetic elements (MGEs). [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • Biogas production from nickel spiked sewage sludge was examined. • Spiking nickel salt had better results on methanation than nano-sized light metal. • 5 mgNi-Salt/kgVS and 2 mg(Nitrilotriacetic acid)/L boosted the biogas yield by 10% • The positive results were achieved in both batch and continuously fed digester. Abstract The presence of micro-nutrients can be stimulatory for the anaerobic digestion (AD) of hardly degradable wastes and thus, improve process performance. Among the essential trace elements, nickel is involved in multiple important enzymes necessary for efficient AD. The present study investigates the effect of nickel spiked sewage sludge on batch and continuous mode operation. Metal spiking was conducted in the form of nanoparticles (Ni-NPs) and salt (NiCl 2 ·6H 2 O). Results from batch assays showed that 5 mgNi-Salt/kgVS in the presence of Nitrilotriacetic acid (NTA) enhanced the methane yield by ∼10% compared to the untreated sample. The impact of Ni-NPs in the AD process was also positive, but slightly lower compared to the effect of NiCl 2 ·6H 2 O. The stimulatory impact of Ni was also revealed in continuously fed digester boosting the methane yield by ∼8%. Overall, the improved methane production indicated that methanogenic archaea were favoured by the simultaneous supplementation of Ni and NTA. [ABSTRACT FROM AUTHOR]

The effects of microwave pretreatment (MW) on co-digestion of food waste (FW) and sewage sludge (SS) have never been investigated. In this study, a series of mesophilic biochemical methane potential (BMP) tests were conducted to determine the optimized ratio of FW and SS based on MW, and the evolution of bacterial and archaeal community was investigated through high-throughput sequencing method. Results showed that the optimized ratio was 3:2 for co-digestion of FW and SS based on MW, and the methane production was 316.24 and 338.44 mLCH 4 /gVS added for MW-FW and MW-SS, respectively. The MW-SS was superior for methane production compared to MW-FW, in which accumulation of propionic acid led to the inhibition of methanogenesis. Proteiniborus and Parabacteroides were responsible for proteins and polysaccharides degradation for all, respectively, while Bacteroides only dominated in co-digestion. Methanosphaera dominated in MW-FW at the active methane production phase, while it was Methanosarcina in MW-SS and mono-SS. [ABSTRACT FROM AUTHOR]

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 bla TEM and mcr-1 , and the phyla of Proteobacteria, Chloroflexi, Synergistetes, Euryarchaeote, intI1 and merA correlated significantly with the fate of bla CTX-M , ereA , tetG and sulI. This study highlighted the importance of substrate types when considering the fate of ARGs during AD. Image 1 • Substrate types significantly influenced the ARGs fate during anaerobic digestion. • The influence from the substrate microbial community on the ARGs fate was limited. • The ARGs in the substrates themselves contributed little to the ARGs fate. • The dominant factors influencing the separate ARGs were determined. • Nutrient variance led to microbial community variance that dominated ARGs fate. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • SRT significantly influenced the ARGs fate in different configurations. • The role of SRT on the ARGs fate varied from ARG types and reactor configurations. • Shorter SRT availed the ARGs reduction for CK and two-stage digestion. • One-stage digestion of pretreated sludge need longer SRT for the ARGs reduction. • Co-selection from heavy metals contributed the most to ARGs fate response to SRT. Abstract 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 CH 4 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. [ABSTRACT FROM AUTHOR]

Swine manure was typical for the combined pollution of heavy metals and antibiotics. The effects of widely used veterinary antibiotic chlortetracycline (CTC), Cu and their combination on swine manure anaerobic digestion performance and microbial community have never been investigated. Thus, four 2 L anaerobic digestion reactors were established including reactor A (control), B (CTC spiked by 0.5 g/kg dry weight, dw), C (Cu spiked by 5 g/kg dw) and D (combination of CTC, 0.5 g/kg dw, and Cu, 5 g/kg dw), and dynamics of bacterial and archaeal community structure was investigated using high throughput sequencing method. Results showed that addition of CTC and Cu separately could increase the total biogas production by 21.6% and 15.8%, respectively, while combination of CTC and Cu severely inhibited anaerobic digestion (by 30.3%). Furthermore, corresponding to different stages and reactors, four kinds of microbes including bacteria and archaea were described in detail, and the effects of CTC, Cu and their combination mainly occurred at hydrolysis and acidification phases. The addition of Cu alone changed the dynamics of archaeal community significantly. It was genus Methanomassiliicoccus that dominated at the active methane production for A, B and D, while it was genus Methanobrevibacter and Methanoculleus for C. [ABSTRACT FROM AUTHOR]

Three anaerobic digestion (AD) processes of waste activated sludge (WAS) were established including the control (mono-WAS), one-stage AD and two-stage AD along with microwave-H 2 O 2 pre-treatment (MW-H 2 O 2 ) to investigate the profiles and drivers of antibiotic resistance genes (ARGs) distribution concerning co-selection from heavy metals, intI1 and microbial community through qPCR and high-throughput sequencing method. Results showed that MW-H 2 O 2 could reduce the absolute gene copies of all ARGs while increased the relative abundance of most ARGs. After subsequent AD, both total ARGs quantities and relative abundance were enriched while two-stage AD showed some advantages over ARGs abundance reduction. Besides, AD was more effective on the potential pathogens reduction than MW-H 2 O 2 . AD could reduce the role of intI1 on the spread of ARGs, while mantel test and procrustes analysis indicated that the variation of ARGs abundance was closely associated with the discrepancy of bacterial community. [ABSTRACT FROM AUTHOR]

As antibiotic and heavy metals are over used in the livestock industry, animal manure is a reservoir of antibiotic resistance genes (ARGs). Anaerobic digestion has been reported to have the potential to reduce ARGs. However, few studies investigated whether reduction of ARGs would be affected by different external pressures including antibiotics and heavy metals during anaerobic digestion. The purpose of this study was thus to investigate effects of both chlortetracycline (CTC) and Cu on reduction of ARGs, heavy metal resistance genes (HMRGs) and mobile genetic elements (MGEs) during the swine manure anaerobic digestion. The results showed that the predominant ARGs ( tet O, tet W, tet X, tet L) could be effectively reduced (approximately 1.00 log copies/g TS) through mesophilic anaerobic digestion. Microbial community evolution was the main driver. It was interesting that Treponema might indicate the termination of anaerobic digestion and compete with ARGs host bacteria. Addition of CTC, Cu and CTC + Cu affected microbial community change and hindered removal of ARGs, especially, CTC + Cu seriously affected Treponema and ARGs during anaerobic digestion. [ABSTRACT FROM AUTHOR]

In this study, anaerobic digestion of mono-SS, MW-SS:FW and SS:MW-FW was investigated to understand the fate of ARGs and its drivers. Anaerobic digestion was effective for the reduction of metal resistance genes (MRGs), and could reduce the abundance of bla OXA-1 , sulI and tetG , while sulII in co-digestion and bla TEM and ereA only in MW-SS. ARGs reduction could be partly attributed to the reduction of co-selective pressure from heavy metals reflected by MRGs. However, the abundance of mefA/E , ermB , ermF , tetM and tetX increased significantly. Anaerobic co-digestion, especially for MW-SS, could reduce total ARGs abundance compared with mono-SS, and evolution of bacterial community was the main driver for the fate of ARGs. [ABSTRACT FROM AUTHOR]

The rheological behavior of sludge is of serious concern in anaerobic digestion. This study investigated the rheological properties of sewage sludge during enhanced anaerobic digestion with microwave-H 2 O 2 pretreatment (MW-H 2 O 2 ). The results showed that MW-H 2 O 2 pretreatment resulted in the improvement of sludge flowability and weakening of its viscoelastic properties. Further positive effects on the rheological properties of digested sludge during anaerobic digestion were observed. The flowability was improved with a low level of apparent viscosity. The decrease of the consistency index and increase of the flow behavior index indicated that the strength of the inner structures and non-Newtonian flow characteristics of digested sludge weakened. Both the storage modulus (G′) and loss modulus (G″) decreased, indicating that the viscoelastic behavior became weak. These effects were possibly attributed to the changes of the digested sludge micro-structures, such as extracellular polymeric substances (EPS). This study concluded that anaerobic digestion for treating sewage sludge combined with pretreatment is a more favorable option than single anaerobic digestion from the perspective of rheology. [ABSTRACT FROM AUTHOR]