Responses of aerobic granular sludge to fluoroquinolones: Microbial community variations, and antibiotic resistance genes. - Related Documents




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804601.0000Responses of aerobic granular sludge to fluoroquinolones: Microbial community variations, and antibiotic resistance genes. In this study, aerobic granular sludge (AGS) was operated under high levels of ammonium for removing three fluoroquinolones (FQs), i.e., ciprofloxacin (CFX), ofloxacin (OFX), and norfloxacin (NFX) at 3, 300, and 900 µg/L, respectively. Two key objectives were to investigate the differential distribution of antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs) in sludge fractions and to evaluate correlations between ARGs and MGEs to nitrifying and denitrifying bacteria. AGS showed excellent stability under the exposure of FQs, with nitrite-oxidizing bacteria (NOB) more sensitive to FQs than ammonium-oxidizing bacteria (AOB). Specific oxygen utilization rates (SOUR) showed a reduction of 26.9% for NOB but only 4.0% of the reduced activity of AOB by 3 μg/L FQs. AGS performed better removal efficiencies for CFX and NFX than OFX, and the efficiencies increased with their elevated concentrations, except at 900 μg/L FQs. The elevated FQ concentrations led to a significant enrichment of intI1 and genus Thauera, while qnrD and qnrS showed no accumulation. Compared to nitrifiers, FQs relevant ARGs and the intI1 gene preferred to exist in denitrifiers, and the abundance of denitrifiers behaved a decreasing trend with the sludge size. Two quinoline-degrading bacteria were found in the AGS system, i.e., Alicycliphilus and Brevundimonas, possibly carrying qnrS and qnrD, respectively. Their relative abundance increased with the sludge size, which was 2.18% in sludge <0.5 mm and increased to 3.70% in sludge >2.0 mm, suggesting that the AGS may be a good choice in treating FQs-containing wastewater.202133676249
804510.9997Correlation among extracellular polymeric substances, tetracycline resistant bacteria and tetracycline resistance genes under trace tetracycline. Antibiotic resistance occurrences and proliferation in activated sludge have attracted more and more attention nowadays. However, the role which extracellular polymeric substance (EPS) plays on the antibiotic resistance is not clear. The changes and correlation among EPS, tetracycline (TC) resistant bacteria (TRB) and TC resistance genes (TRGs) of sequencing batch reactors (SBRs) were investigated. Performance of SBR without TC was compared with two other SBRs to which different amounts of TC were added. Total average EPS contents were found to increase significantly from 66 mg g−1 VSS to 181 mg g−1 VSS as the TC concentrations increased from 0 to 100 μg L−1. As the EPS content increased, TRB in sludge of the three SBRs increased significantly from 105 to 106 colony forming unit mL−1 after being exposed to TC. In addition, the concentrations of three groups of TRGs (copies mL−1) were determined by real-time fluorescence quantitative polymerase chain reaction and followed the order: efflux pump genes > ribosome protected genes > degradation enzyme genes. The numbers of TRGs in the idle stage were larger than those in the aeration sludge. Correlation coefficients (R2) between EPS and TRB in sludge were 0.823 (p < 0.01) while the correlation between EPS and total TRGs was poor (R2 = 0.463, p > 0.05). But it showed the same tendency that EPS and TRGs in sludge increased with the increasing of TC.201425461932
801820.9997Analysis for microbial denitrification and antibiotic resistance during anaerobic digestion of cattle manure containing antibiotic. This study investigated the effects of tylosin (0, 10, and 100 mg/kg dry weight) on the denitrification genes and microbial community during the anaerobic digestion of cattle manure. N(2) emissions were reduced and N(2)O emissions were increased by 10 mg/kg tylosin. Adding 100 mg/kg tylosin increased the emission of both N(2)O and N(2). The different responses of denitrifying bacteria and genes to tylosin may have been due to the presence of antibiotic resistance genes (ARGs). Network analysis indicated that denitrification genes and ARGs had the same potential host bacteria. intI1 was more important for the horizontal transfer of denitrification genes and ARGs during anaerobic digestion than intI2. The anaerobic digestion of manure containing tylosin may increase nitrogen losses and the associated ecological risk.201931326686
801130.9996Co-selective Pressure of Cadmium and Doxycycline on the Antibiotic and Heavy Metal Resistance Genes in Ditch Wetlands. Abuse of heavy metals and antibiotics results in the dissemination of metal resistance genes (MRGs) and antibiotic resistance genes (ARGs). Ditch wetlands are important sinks for heavy metals and antibiotics. The relationships between bacterial communities and MRG/ARG dissemination under dual stresses of heavy metals and antibiotics remain unclear. The responses of MRGs and ARGs to the co-selective pressure of cadmium (Cd) and doxycycline (DC) in ditch wetlands were investigated after 7-day and 84-day exposures. In ecological ditches, residual rates of Cd and DC varied from 0.4 to -5.73% and 0 to -0.61%, respectively. The greatest total relative abundance of ARGs was observed in the Cd 5 mg L(-1) + DC 50 mg L(-1) group. A significant level of DC (50 mg L(-1)) significantly reduced the total relative abundances of MRGs at a concentration of 5 mg L(-1) Cd stress. Redundancy analysis indicated that Cd and DC had strong positive effects on most ARGs and MRGs after a 7-day exposure. Meanwhile, the class 1 integron gene (intI1) exhibited strong positive correlations with most ARGs and cadmium resistance genes (czcA) after an 84-day exposure. Network analysis showed that Acinetobacter and Pseudomonas were the potential dominant host genera for ARGs and MRGs, and tetracycline resistance genes (tetA), czcA, and intI1 shared the same potential host bacteria Trichococcus after an 84-day exposure.202235250936
779540.9996Factors influencing the removal of antibiotic-resistant bacteria and antibiotic resistance genes by the electrokinetic treatment. The performance of the electrokinetic remediation process on the removal of antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs) was evaluated with different influencing factors. With chlortetracycline (CTC), oxytetracycline (OTC), and tetracycline (TC) as template chemicals, the removal of both ARB and ARGs was enhanced with the increase of voltage gradient (0.4-1.2 V cm(-1)) and prolonged reaction time (3-14 d). The greatest removal (26.01-31.48% for ARB, 37.93-83.10% for ARGs) was obtained applying a voltage of 1.2 V cm(-1), leading to the highest electrical consumption. The effect of polarity reversal intervals on the inactivation ratio of ARB followed the order of 0 h (66.06-80.00%) > 12 h (17.07-24.75%) > 24 h (10.44-13.93%). Lower pH, higher current density, and more evenly-distributed voltage drop was observed with a polarity reversal interval of 12 h compared with that of 24 h, leading to more efficient electrochemical reactions in soil. Compared with sul genes, tet genes were more vulnerable to be attacked in an electric field. It was mainly attributed to the lower abundance of tet genes (except tetM) and the varied effects of electrokinetic remediation process on different ARGs. Moreover, a relatively less removal ratio of tetC and tetG was obtained mainly due to the mechanism of the efflux pump upregulation. Both tet and sul genes were positively correlated with TC-resistant bacteria. The efflux pump genes like tetG and the cellular protection genes like tetM showed different correlations with ARB. This study enhances the current understanding on the removal strategies of ARB and ARGs, and it provides important parameters for their destruction by the electrokinetic treatment.201829807293
799650.9996A sludge bulking wastewater treatment plant with an oxidation ditch-denitrification filter in a cold region: bacterial community composition and antibiotic resistance genes. Bacterial community structure of activated sludge directly affects the stable operation of WWTPS, and these bacterial communities may carry a variety of antibiotic resistance genes (ARGs), which is a threat to the public health. This study employed 16S rRNA gene sequencing and metagenomic sequencing to investigate the bacterial community composition and the ARGs in a sludge bulking oxidation ditch-denitrification filter WWTP in a cold region. The results showed that Trichococcus (20.34%), Blautia (7.72%), and Faecalibacterium (3.64%) were the main bacterial genera in the influent. The relative abundances of norank_f_Saprospiraceae and Candidatus_Microthrix reached 10.24% and 8.40%, respectively, in bulking sludge, and those of norank_f_Saprospiraceae and Candidatus_Microthrix decreased to 6.56 and 7.10% after the anaerobic tank, indicating that the anaerobic tank had an inhibitory effect on filamentous bacteria. After 20 mJ/cm(2) UV disinfection, about 540 bacterial genera, such as Romboutsia (7.99%), Rhodoferax (7.98%), and Thermomonas (4.13%), could still be detected in the effluent. The ARGs were 345.11 ppm in the influent and 11.20 ppm in the effluent; 17 subtypes, such as sul1, msrE, aadA5, ErmF, and tet(A), could be detected throughout the entire process. These ARG subtypes were persistent ARGs with a high health risk. Network analysis indicated that the changes in filamentous bacteria norank_f_Saprospiraceae abundance mainly contributed to the abundance shift of MexB, and Acinetobacter mainly increased the abundance of drfA1. These results above will provide theoretical support for the sludge bulking and ARGs controls of WWTPs in cold regions.202336495431
804860.9996Ecological risks of sulfonamides and quinolones degradation intermediates: Toxicity, microbial community, and antibiotic resistance genes. The ecological risks posed by incompletely degraded antibiotic intermediates in aquatic environments warrant significant attention. This study investigated the degradation mechanisms of sulfonamides (sulfadiazine, sulfamethoxazole) and quinolones (ciprofloxacin, norfloxacin) during thermally activated persulfate (TAP) treatment. The main degradation mechanisms for sulfonamides involved S-N bond cleavage and -NH(2) oxidation mediated by sulfate and hydroxyl radicals, whereas quinolone degradation occurred primarily through piperazine ring cleavage facilitated by a single linear oxygen. Toxic degradation intermediates were found to be enriched with bacteria in real water samples, including Aeromonas (SDZ-50, 9.61%), Acinetobacter (SMZ-50, 21.91%), unclassified Archaea (CIP-50, 19.32%), and Herbaspirillum (NOR-50, 17.36%). Meanwhile, the abundance of sulfonamide-associated antibiotic resistance genes (ARGs) (sul1 and sul2) and quinolone-associated ARGs (mfpA, emrA, and lfrA) significantly increased, with SMZ-50 and NOR-50 reaching 659.34 and 2009.98 RPKM, respectively. Correlation analysis revealed differences in host diversity and composition driven by the same classes of antibiotics and their intermediates.202539662843
719270.9996Sulfamethoxazole and COD increase abundance of sulfonamide resistance genes and change bacterial community structures within sequencing batch reactors. The abundant microbial community in biological treatment processes in wastewater treatment plants (WWTPs) may potentially enhance the horizontal gene transfer of antibiotic resistance genes with the presence of antibiotics. A lab-scale sequencing batch reactor was designed to investigate response of sulfonamide resistance genes (sulI, sulII) and bacterial communities to various concentrations of sulfamethoxazole (SMX) and chemical oxygen demand (COD) of wastewater. The SMX concentrations (0.001 mg/L, 0.1 mg/L and 10 mg/L) decreased with treatment time and higher SMX level was more difficult to remove. The presence of SMX also significantly reduced the removal efficiency of ammonia nitrogen, affecting the normal function of WWTPs. All three concentrations of SMX raised both sulI and sulII genes with higher concentrations exhibiting greater increases. The abundance of sul genes was positive correlated with treatment time and followed the second-order reaction kinetic model. Interestingly, these two genes have rather similar activity. SulI and sulII gene abundance also performed similar response to COD. Simpson index and Shannon-Weiner index did not show changes in the microbial community diversity. However, the 16S rRNA gene cloning and sequencing results showed the bacterial community structures varied during different stages. The results demonstrated that influent antibiotics into WWTPs may facilitate selection of ARGs and affect the wastewater conventional treatment as well as the bacteria community structures.201728211331
807280.9996Response of microbial communities based on full-scale classification and antibiotic resistance genes to azithromycin and copper combined pollution in activated sludge nitrification laboratory mesocosms at low temperature. This study aimed to investigate the short-term response of abundant-rare genera and antibiotic resistance genes (ARGs) to azithromycin (AZM, 0.05-40 mg/L) and copper (1 mg/L) combined pollution in activated sludge nitrification system at low temperature. Nitrification was as expected inhibited in stress- and post-effects periods under AZM concentration higher than 5 mg/L. Abundant and rare taxa presented dissimilar responses based on full-scale classification. Conditionally rare or abundant taxa (CRAT) were keystone taxa. Relative abundance of ammonia-oxidizing archaea increased, and three aerobic denitrifying bacteria (Brevundimonas, Comamonas and Trichococcus) were enriched (from 9.83% to 68.91% in total). Ammonia nitrogen assimilating into Org-N and denitrification may be nitrogen pathways based on predict analysis. 29 ARGs were found with more co-occurrence patterns and high concentration of AZM (greater than 5 mg/L) caused their proliferation. Importantly, expect for some abundant taxa, rare taxa, potential pathogens and nitrogen-removal functional genera were the main potential hosts of ARGs.202134523571
804290.9996Algal-bacterial consortium mediated system offers effective removal of nitrogen nutrients and antibiotic resistance genes. The sulfonamide antibiotic resistance genes (ARGs) especially sul1 was identified as the dominant in eutrophic water. The performance of Chlorella vulgaris-B. licheniformis consortium toward sul1 removal, total nitrogen (TN) removal, and the mechanism of sul1 removal was investigated. The removal efficiency of exogenous ARGs plasmids carrying sul1 reached (97.2 ± 2.3)%. The TN removal rate reached (98.5 ± 1.2)%. The enhancements of carbon metabolism, nitrogen metabolism, aminoacyl-tRNA biosynthesis, and glycoproteins had significant influences on sul1 and TN removals, under the premise of normal growth of algae and bacteria. The quantitative polymerase chain reaction (qPCR) results suggested that the absolute abundances of sul1 were low in algal-bacterial systems (0 gene copies/mL) compared with individual systems ((1 × 10(6) ± 15) gene copies/mL). The duplication of sul1 was inhibited in algal cells and bacterial cells. The algal-bacterial consortium seems to be a promising technology for wastewater treatment with a potential to overcome the eutrophication and ARGs challenges.202236049708
8041100.9996Insights into the microalgae-bacteria consortia treating swine wastewater: Symbiotic mechanism and resistance genes analysis. This study investigated the effects of microalgae-bacteria consortia (MBC) (Chlorella pyrenoidosa-activated sludge (AS)) treating swine wastewater with low C/N ratios. After co-culture, the removal rates of NH(4)(+)-N and PO(4)(3-)-P increased by 53.84% and 43.52%. Furthermore, the sulfamethoxazole (SMX) degradation rates in MBC were slightly higher than in the activated sludge process. Interestingly, the absolute abundance of antibiotic resistance genes (ARGs) in effluent from MBC is relatively less than in the AS process. C. pyrenoidosa has a negative zeta potential that allows bacteria to adhere to its surface. The concentrations of carbohydrates and proteins in extracellular polymeric substance (EPS) of MBC dramatically increased compared with the AS process. At the phylum level, Proteobacteria, Bacteroidota, and Cyanobacteria were the main bacteria, while Ascomycota and Basidiomycota were the primary fungi in MBC. Overall, those findings lead to a better understanding of the swine wastewater containing antibiotic treatment by MBC.202235217162
8013110.9996New insight into fates of sulfonamide and tetracycline resistance genes and resistant bacteria during anaerobic digestion of manure at thermophilic and mesophilic temperatures. This study investigated the variations in antibiotic (sulfonamide and tetracycline) resistance genes (ARGs) and resistant bacteria (ARB) during manure anaerobic digestion (AD) at 35 ℃ and 55 ℃, and discussed the mechanisms of variations in ARGs. The AD lasted for 60 days, five ARGs and intI1 each decreased in abundance after AD at the thermophilic temperature, while only half decreased at the mesophilic temperature. On days 10, 30, and 60, sulfonamide and tetracycline ARB were screened on selective media. During thermophilic AD, ARB numbers reduced by 4-log CFUs per gram dry manure, but only by approximately 1-log CFU at the mesophilic temperature. However, ARB composition analysis showed that at either temperature, no significant reduction in identified ARB species was observed. Furthermore, 72 ARB clones were randomly selected to detect the ARGs they harbored, and the results showed that each ARG was harbored by various hosts, and no definitive link existed between ARGs and bacterial species. In addition, by comparison with the identified host by culture method, the host prediction results based on the correlation analysis between ARGs and the bacterial community was proven to be unreliable. Overall, these findings indicated that relationships between ARB and ARGs were intricate.202031685315
8012120.9996Sensitive response mechanism of ARGs and MGEs to initial designed temperature during swine manure and food waste co-composting. The rapid aerobic composting process has been used to reduce organic wastes, but the associated risks of antibiotic resistance genes (ARGs) need to evaluate in an efficient way. The primary objective of this work was to explore the underlying mechanism of initial adjustment in composting temperature on the variation of ARGs, mobile genetic elements (MGEs), and microbial composition during co-composting. The co-composting was initially externally heated (T2) for 5 days. The results showed that ARGs abundance in conventional composting (T1) was reduced by 49.36%, while multidrug was enriched by 86.16% after a period of 30 days. While in T2 ARGs were removed by 79.46% particularly the fraction of sulfonamide, multidrug, and vancomycin resistance genes were >90% without rebounding of any ARGs. Whereas, MGEs were reduced by 68.12% and 93.62% in T1 and T2, while the half-lives of ARGs and MGEs were lower in T2 compared to T1 (86.3%,86.7%). T2 also affected the metabolism function by regulating carbohydrate metabolism (9.62-10.39%) and amino acid metabolism (9.92-10.93%). Apart from this, the potential human pathogenic bacteria Pseudomonas was reduced by 90.6% in T2 and only 32.9% in T1 respectively. Network analysis showed that Ureibacillus, Weissella, Corynebacterium, Escherichia-Shigella, Acinetobacter were the main host of multiple genes. Structural equation models exhibited that bacterial communities were mainly responsible for the enrichment of ARGs in T1, whereas, it was directly affected by MGEs in T2. Similarly, ARGs variation was directly related to composting temperature. With this simple strategy, ARGs associated risk can be significantly reduced in composting.202336208781
7793130.9996Treatment of pharmaceutical wastewater by ionizing radiation: Removal of antibiotics, antimicrobial resistance genes and antimicrobial activity. In present study, the treatment of real pharmaceutical wastewater from an erythromycin (ERY) production factory by gamma irradiation was investigated. Results showed that a variety of antimicrobial resistance genes (ARGs), involving MLSB, tet, bla, multidrug, sul, MGEs and van genes and plentiful 9 bacterial phyla were identified in the raw wastewater. In addition to ERY, sulfamethoxazole (SMX) and tetracycline (TC) were also identified with the concentration of 3 order of magnitude lower than ERY. Results showed that the abatement of ARGs and antibiotics was much higher than that of antimicrobial activity and COD. With the absorbed dose of 50 kGy, the removal percentage of ARGs, ERY, antimicrobial activity and COD was 96.5-99.8%, 90.0%, 47.8% and 10.3%, respectively. The culturable bacteria were abated fast and completely at 5.0 kGy during gamma irradiation. The genus Pseudomonas was predominant in raw wastewater (56.7%) and its relative abundance decreased after gamma irradiation, to 1.3% at 50 kGy. With addition of peroxymonosulfate (PMS, 50 mM), the antimicrobial activity disappeared completely and ERY removal reached as high as 99.2% at the lower absorbed dose of 25 kGy. Ionizing radiation-coupled technique is a potential option to treat pharmaceutical wastewater for reduction of antibiotics, ARGs and antimicrobial activity.202134088196
7960140.9996Diversity evolution of functional bacteria and resistance genes (CzcA) in aerobic activated sludge under Cd(II) stress. An activated sludge sequencing batch reactor (SBR) was used to treat divalent cadmium (Cd(II)) wastewater for 60 d to investigate the overall treatment performance, evolution of the bacterial community, and abundance of the Cd(II) resistance gene CzcA and shifts in its potential host bacteria. During stable operation with a Cd(II) concentration of 20 mg/L, the average removal efficiencies of Cd(II) and chemical oxygen demand (COD) were more than 85% and that of total phosphorus was greater than 70%, while the total nitrogen (TN) was only about 45%. The protein (PN) content in the extracellular polymeric substances (EPS) increased significantly after Cd(II) addition, while polysaccharides displayed a decreasing trend (p < 0.05), indicating that EPS prefer to release PN to adsorb Cd(II) and protect bacteria from damage. Three-dimensional fluorescence spectral analysis showed that fulvic acid-like substances were the most abundant chemical components of EPS. The addition of Cd(II) adversely affected most denitrifying bacteria (p < 0.05), which is consistent with the low TN removal. In addition, quantitative polymerase chain reaction analysis revealed that CzcA gene abundance decreased as the Cd(II) concentration increased, possibly because expression of the CzcA gene was inhibited by Cd(II) stress. The majority of CzcA gene sequences were carried by Pseudomonas, making it the dominant genus among Cd(II)-resistant bacteria.201931514000
8071150.9996Impact of ciprofloxacin and copper combined pollution on activated sludge: Abundant-rare taxa and antibiotic resistance genes. This study aimed to explore the impacts of ciprofloxacin (CIP, 0.05-40 mg/L) and copper (3 mg/L) combined pollution on nitrification, microbial community and antibiotic resistance genes (ARGs) in activated sludge system during stress- and post-effect periods. Higher CIP concentration inhibited nitrification and an average of 50% total nitrogen removal occurred under 40 mg/L of CIP pressure. The stress- and post-effects on bacterial diversity and structure were obviously distinct. Abundant genera were more sensitive to combined pollution than rare genera based on full-scale classification and conditionally rare or abundant taxa were keystone taxa in their interactions. Ammonia oxidation genes were inhibited under high CIP level, but some aerobic denitrifying bacteria (Thauera, Comamonas and Azoarcus) and key genes increased. 96 ARG subtypes were detected with complex positive relationships and their potential hosts (abundant-rare-functional genera) changed in two periods. This study highlights the different stress- and post-effects of combined pollution on activated sludge.202235217161
8019160.9996In-feed antibiotic use changed the behaviors of oxytetracycline, sulfamerazine, and ciprofloxacin and related antibiotic resistance genes during swine manure composting. The dynamics of oxytetracycline (OTC), sulfamerazine (SM1), ciprofloxacin (CIP) and related antibiotic resistance genes (ARGs) during swine manure composting were compared between manure collected from swine fed a diet containing these three antibiotics (T(D)) and manure directly spiked with these drugs (T(S)). The composting removal efficiency of OTC (94.9 %) and CIP (87.8 %) in the T(D) treatment was significantly higher than that of OTC (83.8 %, P <  0.01) and CIP (83.9 %, P <  0.05) in the T(S) treatment, while SM1 exhibited no significant difference (P >  0.05) between the two treatments. Composting effectively reduced the majority of ARGs in both T(D) and T(S) types of manure, especially tetracycline resistance genes (TRGs). Compared with the T(S) treatment, the abundance of some ARGs, such as tetG, qepA, sul1 and sul2, increased dramatically up to 309-fold in the T(D) treatment. The microbial composition of the composting system changed significantly during composting due to antibiotic feeding. Redundancy analysis suggested that the abundance of ARGs had a considerable impact on alterations in the physicochemical parameters (C/N, pH and temperature) and bacterial communities (Actinobacteria, Proteobacteria and Firmicutes) during the composting of swine manure.202133254754
8053170.9996Increasing the removal efficiency of antibiotic resistance through anaerobic digestion with free nitrous acid pretreatment. Swine manure is a significant reservoir for antibiotic resistance. Anaerobic digestion (AD) is a common biological process used to treat swine manure but still faces low efficiencies in biogas production and antibiotic resistance removal. It is here shown that AD with free nitrous acid pretreatment (FNA) was effective in reducing antibiotic resistance genes (ARGs) in swine manure. FNA pretreatment (nitrite =250 mg N/L, pH=5.0, temperature=20 ± 1 °C) simultaneously reduced antibiotics (Tetracyclines, Quinones and Sulfonamides), inactivated antibiotics resistance bacteria (ARB) by 0.5-3 logs, and decreased ARGs tet, sul and qnr by 1-2, 1-3 and 0.5 logs, respectively. In the following AD step, the total residual ARGs was reduced to ~3.49 × 10(7) gene copies/g dry total solids (TS), ~1 log lower than that in the AD without pretreatment (3.55 ×10(8) gene copies/g dry TS). Microbial community and network analyses revealed that the ARG removal was mainly driven by the direct FNA effect on reducing ARGs and antibiotics, not related to ARB. Besides, the FNA pretreatment doubled biochemical methane production potential from swine manure. Together these results demonstrate that AD with FNA pretreatment is a useful process greatly facilitating swine manure management.202235816802
8038180.9996Hydroxyl radicals dominated the reduction of antibiotic resistance genes by inactivating Gram-negative bacteria during soil electrokinetic treatment. Antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs) are emerging contaminants that widely exist in the environment. Effective reduction of ARB and ARGs from soil and water could be achieved by electrokinetic remediation (EKR) technology. In water, hydroxyl radicals (·OH) are proved to play a major role in the EKR process; while the reduction mechanism of ARB and ARGs is still unclear in soil. In this study, different concentrations of hydroxyl radical scavengers (salicylic acid) were added to the EKR system to explore the possible role of ·OH in the reduction of ARB and ARGs. The results showed that generally, ·OH played a more vital role in the reduction of ARB (65.24-72.46%) compared to the reduction of total cultivable bacteria (57.50%). And ·OH contributed to a higher reduction of sul genes (60.94%) compared to tet genes (47.71%) and integrons (36.02%). It was found that the abundance of Gram-negative bacteria (Chloroflexi, Acidobacteria and norank_c_Acidobacteria) was significantly reduced, and the correlation between norank_f_Gemmatimonadaceae and sul1 was weakened in the presence of ·OH. Correlation analysis indicated that the abundance of ARGs (especially sul1) was closely related to the Gram-negative bacteria (Proteobacteria, Acidobacteria, and Gemmatimonadetes) in the soil EKR treatment. Moreover, changes in bacterial community structure affected the abundance of ARB and ARGs indirectly. Overall, this study revealed the reduction mechanism of ARB and ARGs by ·OH in the soil EKR system for the first time. These findings provide valuable support for soil remediation efforts focusing on controlling antibiotic resistance.202439312876
8004190.9996Elimination of antibiotic-resistant bacteria and resistance genes by earthworms during vermifiltration treatment of excess sludge. Vermifiltration (VF) and a conventional biofilter (BF, no earthworm) were investigated by metagenomics to evaluate the removal rates of antibiotic-resistant bacteria (ARB), antibiotic resistance genes (ARGs), and class 1 integron-integrase (intI1), as well as the impact mechanism in combination with the microbial community. According to the findings of qPCR and metagenomics, the VF facilitated greater removal rates of ARGs (78.83% ± 17.37%) and ARB (48.23% ± 2.69%) than the BF (56.33% ± 14.93%, 20.21% ± 6.27%). Compared to the control, the higher biological activity of the VF induced an increase of over 60% in the inhibitory effect of earthworm coelomic fluid on ARB. The removal rates of ARGs by earthworm guts also reached over 22%. In addition, earthworms enhanced the decomposition of refractory organics, toxic, and harmful organics, which led to a lower selective pressure on ARGs and ARB. It provides a strategy for reducing resistant pollution in sewage treatment plants and recognizing the harmless stability of sludge.202438170354