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 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.

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.

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.

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.

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.

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.

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 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.

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.

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.

Metatranscriptome Revealed the Efficacy and Safety of a Prospective Approach for Agricultural Wastewater Reuse: Achieving Ammonia Retention during Biological Treatment by a Novel Natural Inhibitor Epsilon-Poly-l-Lysine.

Appropriate inhibitors might play important roles in achieving ammonia retention in biological wastewater treatment and its reuse in agriculture. In this study, the feasibility of epsilon-poly-l-lysine (ε-PL) as a novel natural ammonia oxidation inhibitor was investigated. Significant inhibition (ammonia oxidation inhibition rate was up to 96.83%) was achieved by treating the sludge with ε-PL (400 mg/L, 12 h soaking) only once and maintaining for six cycles. Meanwhile, the organic matter and nitrite removal was not affected. This method was effective under the common environmental conditions of biological wastewater treatment. Metatranscriptome uncovered the possible action mechanisms of ε-PL. The ammonia oxidation inhibition was due to the co-decrease of Nitrosomonas abundance, ammonia oxidation genes, and the cellular responses of Nitrosomonas. Thauera and Dechloromonas could adapt to ε-PL by stimulating stress responses, which maintained the organic matter and nitrite removal. Importantly, ε-PL did not cause the enhancement of antibiotic resistance genes and virulent factors. Therefore, ε-PL showed a great potential of ammonia retention, which could be applied in the biological treatment of wastewater for agricultural reuse.

Quaternary ammonium compounds promoted anoxic sludge granulation and altered propagation risk of intracellular and extracellular antibiotic resistance genes.

Surfactants could influence sludge morphology and disinfectants were linked to antibiotic resistance genes (ARGs). Thus, the response of activated sludge and ARGs to long-term quaternary ammonium compounds (QACs) exposure required further investigation, which is a popular surfactant and disinfectant. Here, three sequencing batch reactors were fed with 5 mg/L most frequently detected QACs (dodecyl trimethyl ammonium chloride (ATMAC C12), dodecyl benzyl dimethyl ammonium chloride (BAC C12) and didodecyl dimethyl ammonium chloride (DADMAC C12)) for 180 d. The long-term inhibitory effect on denitrification ranked: DADMAC C12 > BAC C12 > ATMAC C12. Besides, obvious granular sludge promoted by the increase of α-Helix/(ß-Sheet + Random coil) appeared in DADMAC C12 system. Moreover, intracellular ARGs increased when denitrification systems encountered QACs acutely but decreased in systems chronically exposed to QACs. Although replication and repair metabolism in ATMAC C12 system was higher, ATMAC C12 significantly promoted proliferation of extracellular ARGs. It was noteworthy that the propagation risk of extracellular ARGs in sludge increased significantly during sludge granulation process, and intracellular sul2 genes in sludge and water both increased with the granular diameter in DADMAC C12 system. The universal utilization of QACs may enhance antibiotic resistance of bacteria in wastewater treatment plants, deserving more attention.

Partial S(0)-driven autotrophic denitrification process facilitated the quick natural enrichment of anammox bacteria at room temperature.

Anaerobic ammonium oxidation (anammox) is well-known to be an environmental and promising biotechnology. However, the natural enrichment of anammox bacteria is still a challenging topic. In this study, partial S(0)-driven autotrophic denitrification (PSAD) was developed to stably supply nitrite, and natural enrichment of anammox bacteria was rapidly realized in a single sequencing moving bed biofilm reactor at room temperature. With the initiation of PSAD, anammox bacteria spontaneously emerged within 12 days, and PSAD-anammox coupling system was realized successfully. And then, the influent concentration of ammonium continuously increased to the same concentration as the nitrate, and the mean total nitrogen removal efficiency reached 92.77 %, which was mainly contributed by anammox. Moreover, the coupling of PSAD and anammox reduced the risk of sulfate emissions. cDNA high throughput sequencing revealed that the relative abundance of Candidatus Brocadia and Thiobacillus reached 39.03 % and 13.48 % at the 88th day. Oligotyping analysis illustrated that GATTTAAT and GTCCCA were the dominant Ca. Brocadia and Thiobacillus oligotypes in PSAD-anammox coupling system, respectively. DNA-based stable isotope probing further deciphered that Thiobacillus was the actual performer of PSAD and supported the nitrite for anammox bacteria in PSAD-anammox coupling system. Overall, this work provided a new strategy to naturally enrich anammox bacteria.

Different acesulfame potassium fate and antibiotic resistance propagation pattern in nitrifying and denitrifying sludge systems.

Acesulfame potassium (ACE-K) is a widely utilized sugar substitute with increasing demand, which is frequently detected in various environmental matrix due to recalcitrance. However, a general consensus on the contribution of nitrifying and denitrifying process to ACE-K removal is lacking. Therefore, ACE-K removal, its effects on antibiotic resistant genes (ARGs) propagation and microbial community in nitrifying sequencing batch reactor (N-SBR) and denitrifying sequencing batch reactor (D-SBR) inoculated with the identical activated sludge were investigated. In this study, ACE-K can be eliminated in N-SBR with satisfying removal efficiency (96.76 ± 8.33 %) after 13 d acclimation, while it remained persistent (average ACE-K removal efficiency of 2.24 ± 1.86 %) in D-SBR during 84 d exposure. Moreover, ACE-K hardly affected the performances of these two types of reactors and had little impact on nitrifying and denitrifying functional genes. However, initial contact with ACE-K would increase ARGs abundance, network analysis showed functional bacteria in each reactor were possible ARGs hosts. Potential ACE-K degrading genera Chelatococcus, Bosea and Aquamicrobium were found in both reactors. LefSe analysis showed that Phyllobacteriaceae containing Aquamicrobium genus was a differentially enriched family in N-SBR. This research might provide a perspective for better understanding factor affecting ACE-K fate in wastewater treatment process and its ecological risks.

Unraveling the impact and mechanism of antipyretic paracetamol on intergenera conjugative plasmid transfer.

Antimicrobial resistance has been considered as a great threat to biosecurity and human health. And the transmission of antibiotic resistance genes (ARGs) by conjugated plasmid is a key factor in the prevalence of antimicrobial resistance. Paracetamol (PRC), one of nonopioid analgesics, is an extensively used antipyretic and mild analgesic worldwide available for numerous prescriptions. It was unclear whether PRC could promote the spread of ARGs. Here, it was demonstrated that PRC promoted intergenera conjugative plasmid transfer in an established conjugation model. Both donor and recipient strains treated by PRC emerged the variations of reactive oxygen species (ROS), SOS response and cell membrane permeability. Correspondingly, transcriptome analysis revealed that the gene expression involved in cell membrane permeability and SOS response was up-regulated significantly after PRC exposure. More directly, PRC also increased the expressions of conjugation related genes of trbG and trbP in donor. This study proved for the first time that PRC could enhance the intergenera conjugative plasmid transfer. Collectively, these findings manifested the potential threat associated with the existence of non-antibiotic substance PRC, which could provide an important insight into antimicrobial resistance spread.

The fate and behavior mechanism of antibiotic resistance genes and microbial communities in flocs, aerobic granular and biofilm sludge under chloroxylenol pressure.

Chloroxylenol (PCMX), an antibacterial agent, has been widely detected in water environment and has toxic effects on biology and ecology. During 270 d, the influence of PCMX on the performance of three nitrification systems was investigated, including floc-based sequencing batch reactor (FSBR), aerobic granule-based SBR (AGSBR) and biofilm SBR (BSBR). The nitrification capability of three systems was inhibited by PCMX, but recovered after domestication, and PCMX made three systems realize partial nitrification for 10, 100 and 35 days, respectively. The extracellular polymeric substances of three systems increased first and then decreased with the increment of PCMX. The granular structure of AGSBR may be conducive to the enrichment of antibiotic resistance genes (ARGs), and almost all ARGs of BSBR were reduced during the addition of 5.0 mg/L PCMX. The microbial community results showed that Rhodococcus as potential degrading bacteria was continuously enriched in three systems. Piscinibacter was regarded as the potential antibiotic resistant bacteria, which was positively associated with multiple ARGs in three systems. Additionally, quaternary ammonium compounds resistance genes had a variety of positive correlations with bacteria in three systems. This study provided a new perspective for the usage and treatment of PCMX.

Distinct bacterial communities and resistance genes enriched by triclocarban-contaminated polyethylene microplastics in antibiotics and heavy metals polluted sewage environment.

Knowledge gaps still surround the question of what biofilms form on contaminated microplastics (MPs) in the antibiotics and (or) heavy metals polluted sewage. In this work, the clean polyethylene microplastics (PE MPs) and triclocarban (TCC)-contaminated PE MPs were cultured in the sewage containing only ampicillin (AMP), only copper (Cu) and both AMP and Cu for 28 days. The results showed that the TCC on PE MPs (with concentration of 2.48 mg/g PE MPs) did not impede the adhesion of the bacteria and the formation of biofilm. Moreover, many potential pathogenic bacteria (Aquabacterium and Pseudoxanthomonas) and potential resistant bacteria (Stenotrophomonas) were more likely to attach on TCC-contaminated PE MPs compared with clean PE MPs. In addition, biofilms of TCC-contaminated PE MPs had highest potential pathogenic functions. TCC-contaminated PE MPs also caused the increases of various resistance genes in both biofilm and sewage. The co-occurrence of TCC, AMP and Cu might exert a stronger selective pressure on bacterial communities and promote the co-selection of resistance genes. In addition, TCC-contaminated PE MPs resulted in higher abundance of five mobile genetic elements (MGEs) (intI1, intI3, tnpA-04, IS613 and trb-C) in sewage, which might further promote the transmission of resistance genes.

Fates of quaternary ammonium compound resistance genes and the corresponding resistant strain in partial nitrification/anammox system under pressure of hexadecyl trimethyl ammonium chloride.

Hexadecyl trimethyl ammonium chloride (ATMAC-C16) is a kind of quaternary ammonium compound (QACs) which is extensively consumed as disinfectants, antimicrobials and surfactants. Here, the partial nitrification/anammox (PN/A) system was exposed to different levels of ATMAC-C16 (0-10 mg/L) and the main objective was to reveal the long-term microbiological responses of PN/A system to ATMAC-C16, importantly, explore the tolerance of PN/A to ATMAC-C16 and the key resistant strain. Nitrogen removal efficiency was influenced by environmental and extreme levels of ATMAC-C16 through mainly affecting the anammox (hzsB) gene. Two types of anammox, Candidatus Jettenia and Candidatus Kuenenia, were enriched under the pressure of ATMAC-C16, which allowed PN/A system to maintain good nitrogen removal performance. ATMAC-C16 might cause the hormesis of entire microbial population in PN/A system, leading to the enhancement of cell viability. ATMAC-C16 decreased the relative abundances of most antibiotics resistance genes (ARGs) but significantly enriched QACs resistance genes (QRGs). The tolerance of PN/A system to ATMAC-C16 might be strengthened by inducing the efflux pumps encoding genes (qacH-01/02). Microbial hosts dynamic and co-selection mechanism among ARGs and QRGs resulted in the opposite trends of qacEdeltal-01/02 and qacH-01/02. Pseudoxanthomonas mexicana was identified as the ATMAC-C16 resistant strain, and its resistance to 10 mg/L ATMAC-C16 might not only obtain by capturing the qacH gene, but also benefit from its own efflux pump system. Therefore, from the perspective of the transmission of resistance genes, especially for QRGs, the spread risk of QRGs and ATMAC-C16 resistant strain in PN/A technique should be taken seriously.