Enhancing Fenton-like reaction mediating performance of covalent organic frameworks through porosity modification.

Covalent organic framework (COF) catalytic photocatalysts mediating Fenton-like reactions have been applied to the treatment of organic dyes in printing and dyeing wastewater. However, the photocatalytic performance of original COF is often unsatisfactory. This study investigated the impact of porosity modification strategies on the performance of COF photocatalysts in mediating the removal of organic dyes via Fenton-like reaction. Porosity modification was achieved by increasing the concentration of acetic acid (HAc) catalyst during COF preparation. The modified TAPB-DMTA COF (12M COF) exhibited excellent adsorption and photocatalytic properties. The Fenton-like reaction mediated by 12M COF photocatalysis removed nearly 96% of malachite green (MG) within 20 min, with a rate constant of 0.091 min-1, which was 2.9 and 6.5 times higher than that of g-C3N4 and original COF under the same reaction conditions, respectively. Additionally, the modulation mechanism of porosity modification on COF photocatalysis was explored. The conduction band (CB) of COF was reduced from -0.14 eV to -0.38 eV after porosity modification, facilitating the generation of longer-lived O2•- in the reaction system, which was conducive to efficient MG removal. Anti-interference experiments showed that the photocatalytic Fenton-like reaction system based on 12 M COF was less affected by common anions, cations and dissolved organics, while maintaining a high MG removal rate in tap water, mid-water, secondary clarifier effluent and river water. In summary, porosity modification was an effective strategy to improve the catalytic performance of original COFs. This study presented an efficient metal-free photocatalyst modification strategy for the Fenton-like reaction while avoiding the production of toxic by-products during dye degradation.

Genomic synergistic efficient carbon fixation and nitrogen removal induced by excessive inorganic carbon in the anammox-centered coupling system.

Given the significance of HCO3- for autotrophic anammox bacteria (AnAOB), excessive HCO3- was always provided in anammox-related systems and engineering applications. However, its impact mechanism on anammox process at genome-level remains unknown. This study firstly established an anammox-centered coupling system that entails heterotrophic partial denitrification (PD) and hydrolytic acidification (A-PDHA) fed mainly with inorganic carbon (high HCO3- concentration and low C/N ratio). Metagenomic binning and metatranscriptomics analyses indicated that high HCO3- concentration enhanced expression of natural most efficient phosphoenolpyruvate (PEP) carboxylase within AnAOB, by up to 30.59 folds. This further induced AnAOB to achieve high-speed carbon-fixing reaction through cross-feeding of phosphate and PEP precursors with heterotrophs. Additionally, the enhanced activity of transporters and catalytic enzymes (up to 4949-fold) induced by low C/N ratio enabled heterotrophs to eliminate extracellular accumulated energy precursors mainly derived from carbon fixation products of AnAOB. This maintained high-speed carbon-fixing reaction within AnAOB and supplemented heterotrophs with organics. Moreover, assimilated energy precursors stimulated nitrogen metabolism enzymes, especially NO2- reductase (968.14 times), in heterotrophs. This established an energy-saving PD-A process mediated by interspecies NO shuttle. These variation resulted in efficient nitrogen removal (>95 %) and reduced external organic carbon demand (67 %) in A-PDHA system. This study unveils the great potential of an anammox-centered autotrophic-heterotrophic coupling system for achieving cost-effective nitrogen removal and enhancing carbon fixation under excessive HCO3- doses.

Whatever does not kill them makes them stronger: Using quaternary ammonia antimicrobials to alleviate the inhibition of ammonia oxidation under perfluorooctanoic acid stress.

Perfluorooctanoic acid (PFOA), benzalkyl dimethylammonium compounds (BAC) and antibiotic resistance genes (ARGs) have negative effects on biological sewage treatment. The performance of nitrification systems under stress of PFOA (0.1-5 mg/L) or/and BAC (0.2-10 mg/L) was explored during 84-day experiments using four sequencing batch reactors, in this study. Low (0.1 mg/L) concentration PFOA had a positive influence on ammonia removal, while medium and high (2 and 5 mg/L) concentrations PFOA caused severe inhibition. Meanwhile, PFOA stress resulted in the enrichment of ARGs in water (w-ARGs). BAC (0-10 mg/L) had no obvious influence on ammonia removal. However, BAC promoted the reduction of ARGs and the bacterial community was the main participator (48.07%) for the spread of ARGs. Interestingly, the joint stress of PFOA and BAC increased the ammonia-oxidizing bacteria (AOB) activity from 5.81 ± 0.19 and 6.05 ± 0.79 mg N/(g MLSS·h) to 7.09 ± 0.87 and 7.23 ± 0.29 mg N/(g MLSS·h) in medium and high concentrations, compared to single stress of PFOA, which was observed for the first time. BAC could reduce bioavailability of PFOA through competitive adsorption and decreasing sludge hydrophobicity by the lower ß-Sheet and α-Helix in tightly bound protein. Furthermore, the joint stress of PFOA and BAC was able to intensify the proliferation of w-ARGs and extracellular ARGs in sludge, and developed the most active horizontal gene transfer mediated by intl1 compared to single stress of PFOA or BAC. The batch tests verified the detoxification capacity of BAC on nitrification under 2.5 mg/L PFOA (48 h exposing), and the maximum alleviation of AOB activity was achieved at BAC and PFOA mass ratio of 2:1. In summary, BAC could be used to alleviate the inhibition of PFOA on ammonia oxidation, providing an efficient and sustainable approach in wastewater treatment.

Hormesis and synergistic effects of disinfectants chloroxylenol and benzethonium chloride on highly efficient heterotrophic nitrification-aerobic denitrification functional strain: From performance to mechanism.

The heterotrophic nitrification-aerobic denitrification (HNAD) strain Exiguobacterium H1 (H1) was isolated in this study. The changes in nitrogen metabolism functions of H1 strain were discussed in presence of disinfectants chloroxylenol (PCMX) and benzethonium chloride (BEC) alone and combined pollution (PCMX+BEC). The H1 strain could use NH4+-N, NO2--N and NO3--N as nitrogen sources and had good nitrogen removal performance under conditions of C/N ratio 25, pH 5-8, 25-35 oC and sodium acetate as carbon. PCMX and BEC alone exhibited hormesis effects on H1 strain which promoted the growth of H1 strain at low concentrations but inhibited it at high concentrations, and combined pollution showed synergistic inhibitory on H1 strain. H1 strain owned a full nitrogen metabolic pathway according to functional genes quantification. PCMX encouraged nitrification process of H1, while BEC and combined pollution mostly blocked nitrogen removal. PCMX, but not BEC, mainly led to the enrichment of resistance genes. These findings will aid in systematic assessment of contaminant tolerance characteristics of HNAD strain and its application prospects.

Effects of polyvinyl chloride microplastics and benzylalkyldimethylethyl compounds on system performance, microbial community and resistance genes in sulfur autotrophic denitrification system.

Benzylalkyldimethylethyl ammonium compounds (BAC) and polyvinyl chloride microplastics (PVC MPs), as the frequently detected pollutants in wastewater treatment plants (WWTPs), have attracted more concerns on their ecosystem risks. Therefore, this study investigated how the sulfur autotrophic denitrification (SAD) system responded to the single and joint stress of PVC MPs (1, 10 and 100 mg/L) and BAC (0.5, 5 and 10 mg/L). After 100 days of operation, the presence of 10 mg/L BAC led to obviously inhibitory effects on system performance and microbial metabolic activity. And the additions of PVC MPs or/and BAC stimulated the proliferation of intracellular resistance genes (RGs), whereas exposure to BAC increased the abundances of extracellular RGs and free RGs in water more significantly. Compared to the joint stress, BAC single stress resulted in higher abundances of free RGs in water, which further increased the risk of RGs propagation. Moreover, the interaction between mobile genetic elements and extracellular polymeric substances further increased the spread of RGs. Pathogens might be the potential hosts of RGs and enriched in SAD system and plastisphere, thereby leading to more serious ecological risks. This study will broaden the understanding of the environmental hazards posed by PVC MPs and BAC in WWTPs.

Natural pyrite and ascorbic acid co-enhance periodate activation for inactivation of antibiotic resistant bacteria and inhibition of resistance genes transmission: A green disinfection process dominated by singlet oxygen.

The transmission of antibiotic resistance genes (ARGs) and the propagation of antibiotic resistant bacteria (ARB) threaten public health security and human health, and greener and more efficient disinfection technologies are expected to be discovered for wastewater treatment. In this study, natural pyrite and ascorbic acid (AA) were proposed as environmental-friendly activator and reductant for periodate (PI) activation to inactivate ARB. The disinfection treatment of PI/pyrite/AA system could inactivate 5.62 log ARB within 30 min, and the lower pH and higher PI and natural pyrite dosage could further boost the disinfection efficiency. The 1O2 and SO4•- were demonstrated to be crucial for the inactivation of ARB in PI/pyrite/AA system. The disinfection process destroyed the morphological structure of ARB, inducing oxidative stress and stimulating the antioxidant system. The PI/pyrite/AA system effectively reduced the intracellular and extracellular DNA concentration and ARGs abundance, inhibiting the propagation of ARGs. The presence of AA facilitated the activation of PI with natural pyrite and significantly increased the concentration of Fe2+ in solution. The reusability of natural pyrite, the safety of the disinfection by-products and the inhibition of ARB regeneration indicated the application potential of PI/pyrite/AA system in wastewater disinfection.

Exposure to zinc and dialkyldimethyl ammonium compound alters bacterial community structure and resistance gene levels in partial sulfur autotrophic denitrification coupled with the Anammox process.

Dialkyldimethyl ammonium compound (DADMAC) is widely used in daily life as a typical disinfectant and often co-exists with the heavy metal zinc in sewage environments. This study investigated the effects of co-exposure to zinc (1 mg/L) and DADMAC (0.2-5 mg/L) on the performance, bacterial community, and resistance genes (RGs) in a partial sulfur autotrophic denitrification coupled with anaerobic ammonium oxidation (PSAD-Anammox) system in a sequencing batch moving bed biofilm reactor for 150 days. Co-exposure to zinc and low concentration (0.2 mg/L) DADMAC did not affect the nitrogen removal ability of the PASD-Anammox system, but increased the abundance and transmission risk of free RGs in water. Co-exposure to zinc and medium-to-high (2-5 mg/L) DADMAC led to fluctuations in and inhibition of nitrogen removal, which might be related to the enrichment of heterotrophic denitrifying bacteria dominated by Denitratisoma. Co-exposure to zinc and high concentration DADMAC (5 mg/L) stimulated the secretion of extracellular polymeric substances and increased the proliferation risk of intracellular RGs in sludge. This study provided insights into the application of PSAD-Anammox system and the ecological risks of wastewater containing zinc and DADMAC.

Phenacetin promoted the rapid start-up and stable maintenance of partial nitrification: Responses of nitrifiers and antibiotic resistance genes.

Phenacetin (PNCT) belongs to one of the earliest synthetic antipyretics. However, impact of PNCT on nitrifying microorganisms in wastewater treatment plants and its potential microbial mechanism was still unclear. In this study, PN could be initiated within six days by PNCT anaerobic soaking treatment (8 mg/L). In order to improve the stable performance of PN, 21 times of PNCT aerobic soaking treatment every three days was conducted and PN was stabilized for 191 days. After PN was damaged, ten times of PNCT aerobic soaking treatment every three days was conducted and PN was recovered after once soaking, maintained over 88 days. Ammonia oxidizing bacteria might change the dominant oligotype to gradually adjust to PNCT, and the increase of abundance and activity of Nitrosomonas promoted the initiation of PN. For nitrite-oxidizing bacteria (NOB), the increase of Candidatus Nitrotoga and Nitrospira destroyed PN, but PN could be recovered after once aerobic soaking illustrating NOB was not resistant to PNCT. KEGG and COG analysis suggested PNCT might disrupt rTCA cycle of Nitrospira, resulting in the decrease of relative abundance of Nitrospira. Moreover, PNCT did not lead to the sharp increase of absolute abundances of antibiotic resistance genes (ARGs), and the risk of ARGs transmission was negligible.

The combination of ciprofloxacin and dialkyldimethyl ammonium compound synergistically proliferated intracellular resistance genes in nitrifying system.

Antibiotics and quaternary ammonium compounds (QACs) usually co-exist in wastewater treatment plants. Hence, three sequencing batch reactors were established and named as R1, R2 and R3, to investigate the effects of individual and combined exposure of different concentrations of ciprofloxacin (CIP) (0.2, 1.0 and 2.0 mg/L) and dialkyldimethyl ammonium compound (DADMAC) (0.4, 2.0 and 4.0 mg/L) on the performance, microbial community structures and resistance genes (RGs) in nitrifying system during 150 days. Results showed that CIP had a slight effect on ammonia oxidation activity, while 2.0 and 4.0 mg/L DADAMAC could obviously inhibit it, and the combination of CIP and DADMAC had a synergistic inhibitory effect. Besides, both CIP and DADMAC caused partial nitrification, and the order of nitrite accumulation rate was ranked as R3 > R2 > R1. The combination of CIP and DADMAC had an antagonistic effect on the increase of sludge particle size and α-Helix/(ß-Sheet + Random coil) was lowest in R3 (0.40). The combination of CIP and DADMAC synergistically stimulated most intracellular RGs in sludge, and the relative abundances of target RGs (e.g., qacEdelta1-01, qacH-01 and qnrS) at the end of operation in R3 were increased by 4.61-18.19 folds compared with those in CK, which were 1.34-5.57 folds higher than the R1 and R2. Moreover, the combination of CIP and DADMAC also promoted the transfer of RGs from sludge to water and enriched more potential hosts of RGs, further promoting the spread of RGs in nitrifying system. Thus, the combined pollution of CIP and DADMAC in wastewaters should attract more attentions.

Mechanisms on the removal of gram-negative/positive antibiotic resistant bacteria and inhibition of horizontal gene transfer by ferrate coupled with peroxydisulfate or peroxymonosulfate.

The existence of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) has been a global public environment and health issue. Due to the different cell structures, gram-positive/negative ARB exhibit various inactivation mechanisms in water disinfection. In this study, a gram-negative ARB Escherichia coli DH5α (E. coli DH5α) was used as a horizontal gene transfer (HGT) donor, while a gram-positive ARB Bacillus as a recipient. To develop an efficient and engineering applicable method in water disinfection, ARB and ARGs removal efficiency of Fe(VI) coupled peroxydisulfate (PDS) or peroxymonosulfate (PMS) was compared, wherein hydroxylamine (HA) was added as a reducing agent. The results indicated that Fe(VI)/PMS/HA showed higher disinfection efficiency than Fe(VI)/PDS/HA. When the concentration of each Fe(VI), PMS, HA was 0.48 mM, 5.15 log E. coli DH5α and 3.57 log Bacillus lost cultivability, while the proportion of recovered cells was 0.0017 % and 0.0566 %, respectively, and HGT was blocked. Intracellular tetA was reduced by 2.49 log. Fe(IV) and/or Fe(V) were proved to be the decisive reactive species. Due to the superiority of low cost as well as high efficiency and practicality, Fe(VI)/PMS/HA has significant application potential in ARB, ARGs removal and HGT inhibition, offering a new insight for wastewater treatment.

The synergistic effect of periodate/ferrate (VI) system on disinfection of antibiotic resistant bacteria and removal of antibiotic resistant genes: The dominance of Fe (IV)/Fe (V).

The proliferation of antibiotic resistant genes (ARGs) and antibiotic resistant bacteria (ARB) caused by antibiotic abuse has raised concerns about the global infectious-disease crisis. This study employed periodate (PI)/ferrate (VI) (Fe (VI)) system to disinfect Gram-negative ARB (Escherichia coli DH5α) and Gram-positive bacteria (Bacillus subtilis ATCC6633). The PI/Fe (VI) system could inactivate 1 × 108 CFU/mL of Gram-negative ARB and Gram-positive bacteria by 4.0 and 2.8 log in 30 min. Neutral and acidic pH, increase of PI dosage and Fe (VI) dosage had positive impacts on the inactivation efficiency of ARB, while alkaline solution and the coexistence of 10 mM Cl-, NO3-, SO42- and 20 mg/L humic acid had slightly negative impacts. The reactive species generated by PI/Fe (VI) system could disrupt the integrity of cell membrane and wall, leading to oxidative stress and lipid peroxidation. Intracellular hereditary substance, including DNA and ARGs (tetA), would leak into the external environment through damaged cells and be degraded. The electron spin resonance analysis and quenching experiments indicated that Fe (IV)/Fe (V) played a leading role in disinfection. Meanwhile, PI/Fe (VI) system also had an efficient removal effect on sulfadiazine, which was expected to inhibit the ARGs transmission from the source.

NH2-MIL-101(Fe)-mediated photo-Fenton reaction enhanced simultaneous removal of nitrogen and refractory organics in anammox process through interfacial electron transfer.

In this study, H2O2 (0.1 ‰) and NH2-MIL-101(Fe)-driven (150 mg/L) photo-Fenton-coupled anammox were proposed to simultaneously improve the removal efficiency of nitrogen and humic acid. Long-term experiments showed that the total nitrogen removal efficiency was increased by the photo-Fenton reaction to 91.9 ± 1.5 % by altering the bioavailability of refractory organics. Correspondingly, the total organic carbon removal efficiency was significantly increased. Microbial community analyses indicated that Candidatus_Brocadia maintained high activity during photo-Fenton reaction and was the most abundant genus in the reactor. Dissimilatory nitrate reduction to ammonium process and denitrification process were enhanced, resulting in reduced NO3--N production. The establishment of electron transfer between microorganisms and NH2-MIL-101 (Fe) improved the charge separation efficiency of the quantum dots and increased the intracellular adenosine triphosphate content of anammox bacteria. These results indicated that photo-Fenton-anammox process promoted the removal of nitrogen and refractory organics in one reactor which had good economic value and application prospects.

Efficient peroxymonosulfate activation by nanoscale zerovalent iron for removal of sulfadiazine and sulfadiazine resistance bacteria: Sulfidated modification or not.

Whether it's necessary to extra chemical synthesis steps to modify nZVI in peroxymonosulfate (PMS) activation process are worth to further investigation. The 56 mg/L nZVI/153.65 mg/L PMS and 56 mg/L sulfidated nZVI (S-nZVI) (S/Fe molar ratio = 1:5)/153.65 mg/L PMS) processes could effectively attain 97.7% (with kobs of 3.7817 min-1) and 97.0% (with kobs of 3.4966 min-1) of the degradation of 20 mg/L sulfadiazine (SDZ) in 1 min, respectively. The nZVI/PMS system could quickly achieve 85.5% degradation of 20 mg/L SDZ in 1 min and effectively inactivate 99.99% of coexisting Pseudomonas. HLS-6 (5.81-log) in 30 min. Electron paramagnetic resonance tests and radical quenching experiments determined SO4•-, HO•, 1O2 and O2•- were responsible for SDZ degradation. The nZVI/PMS system could still achieve the satisfactory degradation efficiency of SDZ under the influence of humic acid (exceeded 96.1%), common anions (exceeded 67.3%), synthetic wastewater effluent (exceeded 90.7%) and real wastewater effluent (exceeded 78.7%). The high degradation efficiency of tetracycline (exceeded 98.9%) and five common disinfectants (exceeded 96.3%) confirmed the applicability of the two systems for pollutants removal. It's no necessary to extra chemical synthesis steps to modify nZVI for PMS activation to remove both chemical and biological pollutants.

Successive exposure to traditional disinfectants and quaternary ammonium compounds enhances resistance genes expression and co-selection in biofilm-based partial nitrification-anammox systems.

Quaternary ammonium compounds (QACs) are recommended disinfectants with surfactant properties, surpassing triclosan (TCS) and chloroxylenol (PCMX). Given the transition from traditional disinfectants, it is essential to investigate their impacts on biological nitrogen removal systems and the fate of resistance genes (RGs). In this study, three biofilm-based partial nitrification-anammox (PN/A) systems were established. A reactor named PD was successively exposed to 1 mg/L PCMX and 3 mg/L dioctadecyldimethylammonium chloride (DODMAC, a common QACs). A reactor named TD was successively exposed to 1 mg/L TCS and 3 mg/L DODMAC. A reactor named CD served as a control with only 3 mg/L DODMAC exposure. Results indicated that the total nitrogen removal performance of CD deteriorated markedly with DODMAC exposure compared to that of PD and TD. This phenomenon correlated closely with variations in RGs and their co-selection patterns. Pre-exposure to PCMX or TCS increased the abundance of RGs in the extracellular DNA of the PN/A biofilm, but reduced RGs abundances in the extracellular DNA of water. The tolerance of the PN/A system to successive exposure to the two disinfectants may be strengthened through co-selection of QACs RGs (qacEdelta1-01, qacEdelta1-02, qacH-01 and qacH-02) and mobile genetic elements (intI1 and tnpA-04). Furthermore, potential hosts of RGs are crucial for maintaining PN/A performance. Accumulation of extracellular polymeric substances, reactive oxygen species, and lactate dehydrogenase plays vital roles in the accumulation and transmission of RGs within the PN/A system.

Fates of extracellular and intracellular antibiotic resistance genes in activated sludge and plastisphere under sulfadiazine pressure.

Microplastics, antibiotics, and antibiotic resistance genes (ARGs) represent prominent emerging contaminants that can potentially hinder the efficacy of biological wastewater treatment and pose health risks. Plastisphere as a distinct ecological niche for microorganisms, acts as a repository for ARGs and potential pathogenic bacteria. Nonetheless, the spread pattern of extracellular ARGs (eARGs) and intracellular ARGs (iARGs) in plastisphere under antibiotic exposure was not yet known. This study aimed to investigate disparities in extracellular polymeric substances (EPS) production, extracellular and intracellular microbial community structures, as well as the transmission of eARGs and iARGs between activated sludge and plastisphere in an anaerobic/anoxic/oxic system under sulfadiazine (SDZ) exposure. SDZ was found to enhance EPS production in activated sludge and plastisphere. Interestingly, as SDZ removal efficiency increased, EPS content decreased in activated sludge and plastisphere collected from oxic zone, and continued to increase in plastisphere samples collected from anaerobic and anoxic zones. There were significant differences in microbial community structure between activated sludge and plastisphere, and the DNA fragments of potential pathogenic bacteria were detected in extracellular samples. SDZ exhibited a promoting effect on the propagation of eARGs, which were more abundant in the plastisphere than in activated sludge, thus heightening the risk of ARGs dissemination. Extracellular mobile genetic elements played a pivotal role in driving the spread of eARGs, while the microbial community induced the changes of iARGs. Potential pathogenic bacteria emerged as potential hosts for ARGs and mobile genetic elements within activated sludge and plastisphere, leading to more serious environmental threats.

Deciphering the interaction impacts between antiseptic benzethonium chloride and biofilm nitrification system: Performance, resistance mechanisms and biodegradation.

Benzethonium chloride (BEC) is one of emerging bacteriostatic agents. BEC-bearing wastewater generated during sanitary applications in food and medication is easily combined with other wastewater streams to flow into wastewater treatment plants. This study focused on the long-term (231 days) impacts of BEC on the sequencing moving bed biofilm nitrification system. Nitrification performance was tolerant to low concentration of BEC (≤ 0.2 mg/L), but the nitrite oxidation was severely inhibited when the concentration of BEC was 1.0-2.0 mg/L. Partial nitrification maintained about 140 days with nitrite accumulation ratio over 80%, mainly caused by the inhibition of Nitrospira, Nitrotoga and Comammox. Notably, BEC exposure in the system might cause the co-selection of antibiotic resistance genes (ARGs) and disinfectant resistance genes (DRGs), and the resistance of biofilm system to BEC was strengthened by efflux pumps mechanism (qacEdelta1 and qacH) and antibiotic deactivation mechanism (aadA, aac(6')-Ib and blaTEM). Extracellular polymeric substances secretion and BEC biodegradation were also contributed to the system microorganisms resisting BEC exposure. In addition, Klebsiella, Enterobacter, Citrobacter and Pseudomonas were isolated and identified as BEC degrading bacteria. The metabolites of N,N-dimethylbenzylamine, N-benzylmethylamine and benzoic acid were identified, and the biodegradation pathway of BEC was proposed. This study brought new knowledge about the fate of BEC in biological treatment units and laid a foundation for its elimination from wastewater.

The knock-on effects of different wastewater feeding modes: Change in microbial communities versus resistance genes in pilot-scale aerobic sludge granulation reactors.

To explore the effects of wastewater feeding modes on the formation of aerobic granular sludge (AGS) and the complex relationships between resistance genes and bacteria, two pilot-scale sequencing batch reactors (SBRs) were established. The SBR with influent wastewater introduced uniformly through pipes at bottom was designated as BSBR, and the SBR with inlet wastewater flowing directly from top was TSBR. BSBR formed dense AGS due to uniform wastewater feeding at bottom, while TSBR failed to cultivate AGS. Metagenomic sequencing illustrated that rapid growth of AGS in BSBR was accompanied with increase of antibiotic resistance genes (ARGs) abundance, but ARGs diminished when the size of AGS was stable. The ARGs continued to elevate in TSBR, and abundance of metal resistance genes (MRGs) was always higher than that in BSBR. Two reactors had markedly different bacterial community, microbes in BSBR owned stronger activity, conferred greater potential to proliferate. AdeF in two systems had the most complex gene-bacteria relationships which would undergo HGT within bacterial genus. The different feeding modes of wastewater directly led to the changing size of sludge, which caused knock-on effects of variations in the abundance of microbial communities and resistance genes. This study provided promising suggestions for the rapid cultivation of AGS and control of resistance genes at pilot-scale.

The responses of microbial metabolic activity, bacterial community and resistance genes under the coexistence of nanoplastics and quaternary ammonium compounds in the sewage environment.

Nanoplastics (NPs) and quaternary ammonium compounds (QACs) are frequently detected in sewage. However, little is known about the risks of coexistence of NPs and QACs. In this study, the responses of microbial metabolic activity, bacterial community and resistance genes (RGs) to the exposure of polyethylene (PE), polylactic acid (PLA), silicon dioxide (SiO2) and dodecyl dimethyl benzyl ammonium chloride (DDBAC) were focused on 2nd and 30th day of incubation in sewer environment. Bacterial community contributed 25.01 % to shape RGs and mobile genetic elements (MGEs) after two days of incubation in sewage and plastisphere. After 30 days of incubation, the most important individual factor (35.82 %) was turned to microbial metabolic activity. The metabolic capacity of the microbial communities in plastisphere was stronger than that from SiO2 samples. Moreover, DDBAC inhibited the metabolic capacity of microorganisms in sewage samples, and increased the absolute abundances of 16S rRNA in plastisphere and sewage samples which might be similar to the hormesis effect. After 30 days of incubation, Aquabacterium was the predominant genus in plastisphere. As for SiO2 samples, Brevundimonas was the predominant genus. QACs RGs (qacEdelta1-01, qacEdelta1-02) and antibiotic RGs (ARGs) (aac(6')-Ib, tetG-1) significantly enriched in plastisphere. There was also co-selection among qacEdelta1-01, qacEdelta1-02 and ARGs. In addition, VadinBC27 which enriched in plastisphere of PLA NPs was positively correlated with the potentially disease-causing genus Pseudomonas. It showed that after 30 days of incubation, plastisphere had an important effect on distribution and transfer of pathogenic bacteria and RGs. Plastisphere of PLA NPs also carried the risk of spreading disease.

The spread of different resistance genes fractions in nitrification system under chronic exposure to varying alkyl chain length benzalkyl dimethylammonium compounds.

Benzalkyl dimethylammonium compounds (BACs) are generally applied as surfactants and disinfectants. In this study, the nitrification systems were exposed to different alkyl chain lengths (C12-C16) and different levels of BACs (0-5 mg/L), respectively, totally 120 days and to explore the chronic effect of BACs on resistance genes (RGs). RGs were classified into four fractions based on activated sludge properties. Ammonia oxidation performance were not significantly affected by BACs, whereas BACs increased the absolute abundance of most intracellular RGs in sludge (si-RGs). Under the exposure of BACs, extracellular RGs in water (we-RGs) showed a decrease trend and si-RGs tended to be converted to we-RGs. Tightly bound-Tyrosine side chain was significantly correlated with most we-RGs, and we-intI1 might contribute to the propagation of RGs. Therefore, the risk of transmission of different fractions of RGs in the nitrification system under the stress of BACs should be taken seriously.

Mitigating the inhibition of antibacterial agent chloroxylenol on nitrification system-The role of Rhodococcus ruber in a bioaugmentation system.

The chloroxylenol (PCMX) degrading strain was successfully isolated from sludge and identified as Rhodococcus ruber (R. ruber). Afterwards, a bioaugmentation system was constructed by seeding R. ruber into nitrifying sludge to fasten degradation efficiency of highly toxic PCMX from wastewater. Results showed that R. ruber presented high PCMX-degrading performance under aerobic conditions, 25 °C, pH 7.0 and inoculum sizes of 4% (v/v). These optimized conditions were used in subsequent bioaugmentation experiment. In bioaugmentation system, R. ruber could detoxify nitrifiers by degrading PCMX, and the content of polysaccharide in extracellular polymeric substances increased. The quantitative polymerase chain reaction results exhibited that the absolute abundance of 16S rRNA gene and ammonia oxidizing bacteria (AOB) slightly elevated in bioaugmentation system. After analyzing the results of high-throughput sequencing, it was found that the loaded R. ruber can colonize successfully and turn into dominant strains in sludge system. Molecular docking simulation showed that PCMX had a weaker suppressed effect on AOB than nitrite oxidizing bacteria, and R. ruber can alleviate the adverse effect. This study could provide a novel strategy for potential application in reinforcement of PCMX removal in wastewater treatment.