Multidrug-resistant plasmid RP4 increases NO and N(2)O yields via the electron transport system in Nitrosomonas europaea ammonia oxidation. - Related Documents




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787901.0000Multidrug-resistant plasmid RP4 increases NO and N(2)O yields via the electron transport system in Nitrosomonas europaea ammonia oxidation. Antibiotic resistance genes (ARGs) have recently become an important public health problem and therefore several studies have characterized ARG composition and distribution. However, few studies have assessed their impact on important functional microorganisms in the environment. Therefore, our study sought to investigate the mechanisms through which multidrug-resistant plasmid RP4 affected the ammonia oxidation capacity of ammonia-oxidizing bacteria, which play a key role in the nitrogen cycle. The ammonia oxidation capacity of N. europaea ATCC25978 (RP4) was significantly inhibited, and NO and N(2)O were produced instead of nitrite. Our findings demonstrated that the decrease in electrons from NH(2)OH decreased the ammonia monooxygenase (AMO) activity, leading to a decrease in ammonia consumption. In the ammonia oxidation process, N. europaea ATCC25978 (RP4) exhibited ATP and NADH accumulation. The corresponding mechanism was the overactivation of Complex Ⅰ, ATPase, and the TCA cycle by the RP4 plasmid. The genes encoding TCA cycle enzymes related to energy generation, including gltA, icd, sucD, and NE0773, were upregulated in N. europaea ATCC25978 (RP4). These results demonstrate the ecological risks of ARGs, including the inhibition of the ammonia oxidation process and an increased production of greenhouse gases such as NO and N(2)O.202337421866
787810.9994Horizontal transfer of the multidrug resistance plasmid RP4 inhibits ammonia nitrogen removal dominated by ammonia-oxidizing bacteria. Antibiotic resistance genes (ARGs) have become an important public health concern. Particularly, although several ARGs have been identified in wastewater treatment plants (WWTPs), very few studies have characterized their impacts on reactor performance. Therefore, our study sought to investigate the effect of a representative conjugative transfer plasmid (RP4) encoding multidrug resistance genes on ammonia oxidation. To achieve this, we established sequencing batch reactors (SBRs) and a conjugation model with E. coli donor strains carrying the RP4 plasmid and a typical ammonia-oxidating (AOB) bacterial strain (Nitrosomonas europaea ATCC 25978) as a recipient to investigate the effect of conjugative transfer of plasmid RP4 on AOB. Our findings demonstrated that the RP4 plasmid carried by the donor strains could be transferred to AOB in the SBR and to Nitrosomonas europaea ATCC 25978. In SBR treated with donor strains carrying the RP4 plasmid, ammonia removal efficiency continuously decreased to 71%. Once the RP4 plasmid entered N. europaea ATCC 25978 in the conjugation model, ammonia removal was significantly inhibited and nitrite generation was decreased. Furthermore, the expression of several functional genes related to ammonia oxidation in AOB was suppressed following the transfer of the RP4 plasmid, including amoA, amoC, hao, nirK, and norB. In contrast, the cytL gene encoding cytochrome P460 was upregulated. These results demonstrated the ecological risk of ARGs in WWTPs, and therefore measures must be taken to avoid their transfer.202235427829
848620.9993Multidrug-resistant plasmid modulates ammonia oxidation efficiency in Nitrosomonas europaea through cyclic di-guanylate and acyl-homoserine lactones pathways. Antibiotic resistance genes present a major public health challenge and have potential implications for global biogeochemical cycles. However, their impacts on biological nitrogen removal systems remain poorly understood. In the ammonia-oxidizing bacteria Nitrosomonas europaea ATCC 19718 harboring the multidrug-resistant plasmid RP4, a significant decrease in ammonia oxidation efficiency was observed, accompanied by markedly elevated levels of cyclic di-guanylate (c-di-GMP) and acyl-homoserine lactones (AHLs), compared to plasmid-free controls. The results demonstrated that c-di-GMP facilitates the secretion of AHLs, while elevated levels of AHLs inhibit the ammonia oxidation efficiency of Nitrosomonas europaea ATCC 19718. These results revealed that RP4 plasmid significantly impaired ammonia oxidation efficiency through the c-di-GMP and AHLs pathways. Our findings indicate that the multidrug-resistant plasmid RP4 adversely affects the nitrogen metabolism of ammonia-oxidizing bacteria, potentially disrupting the nitrogen biogeochemical cycle and posing substantial ecological and environmental risks.202640945801
796930.9993Metagenomic insights into the influence of pH on antibiotic removal and antibiotic resistance during nitritation: Regulations on functional genus and genes. The changes in pH and the resulting presence of free nitrous acid (FNA) or free ammonia (FA) often inhibit antibiotic biodegradation during nitritation. However, the specific mechanisms through which pH, FNA and FA influence antibiotic removal and the fate of antibiotic resistance genes (ARGs) are not yet fully understood. In this study, the effects of pH, FNA, and FA on the removal of cefalexin and amoxicillin during nitritation were investigated. The results revealed that the decreased antibiotic removal under both acidic condition (pH 4.5) and alkaline condition (pH 9.5) was due to the inhibition of the expression of amoA in ammonia-oxidizing bacteria and functional genes (hydrolase-encoding genes, transferase-encoding genes, lyase-encoding genes, and oxidoreductase-encoding genes) in heterotrophs. Furthermore, acidity was the primary inhibitor of antibiotic removal at pH 4.5, followed by FNA. Antibiotic removal was primarily inhibited by alkalinity at pH 9.5, followed by FA. The proliferation of ARGs mediated by mobile genetic element was promoted under both acidic and alkaline conditions, attributed to the promotion of FNA and FA, respectively. Overall, this study highlights the inhibitory effects of acidity and alkalinity on antibiotic removal during nitritation.202439068965
850840.9992Phenolic compounds promote the horizontal transfer of antibiotic resistance genes in activated sludge. Phenolic compounds are common organic pollutants in wastewater. During the wastewater treatment process, these compounds may influence the microbial community structure and functions. However, the impact of the phenolic compounds in the wastewater treatment plants on the horizontal transfer of antibiotic resistance genes (ARGs) has not been well assessed. In this study, we investigated the horizontal transfer of ARGs under the stress of phenolic compounds. The results showed that in pure culture bacteria system, p-nitrophenol (PNP), p-aminophenol (PAP) and phenol (PhOH) (10-100 mg/L) can significantly increase the horizontal transfer frequency of ARGs by 2.2-4.6, 3.6-9.4 and 1.9-9.0 fold, respectively. And, the RP4 plasmid transfer from Escherichia coli HB101 (E. coli HB101) to the bacteria in activated sludge increased obviously under the stress of phenolic compounds. Further investigation revealed that the PNP and PhOH at the concentration of 10-100 mg/L increased the production of reactive oxygen species and the permeability of cell membrane in the donor and recipient, which could be the causes of horizontal transfer of RP4 plasmid. In addition, it was also found that PNP, PAP and PhOH stress inhibit the expression of the global regulatory genes korB and trbA in the RP4 plasmid, and increase the expression level of the traF gene, thereby promoting the conjugative transfer of the RP4 plasmid. Taken together, these results improved our understanding of the horizontal transfer of ARGs under the stress of phenolic compounds and provided basic information for management of the systems that treat wastewater containing phenolic compounds.202134392203
795850.9992Microbial response and recovery strategy of the anammox process under ciprofloxacin stress from pure strain and consortia perspectives. Ciprofloxacin (CIP) poses a high risk of resistance development in water environments. Therefore, comprehensive effects and recovery strategies of CIP in anaerobic ammonia oxidation (anammox) process were systematically elucidated from consortia and pure strains perspectives. The anammox consortia was not significantly affected by the stress of 10 mg L(-1) CIP, while the higher concentration (20 mg L(-1)) of CIP caused a dramatic reduction in the nitrogen removal performance of anammox system. Simultaneously, the abundances of dominant functional bacteria and corresponding genes also significantly decreased. Such inhibition could not be mitigated by the recovery strategy of adding hydrazine and hydroxylamine. Reducing nitrogen load rate from 5.1 to 1.4 kg N m(-3) d(-)(1) promoted the restoration of three reactors. In addition, the robustness and recovery of anammox systems was evaluated using starvation and shock strategies. Simultaneously, antibiotic resistance genes and key metabolic pathways of anammox consortia were upregulated, such as carbohydrate and energy metabolisms. In addition, 11 pure stains were isolated from the anammox system and identified through phylogenetic analysis, 40 % of which showed multidrug resistance, especially Pseudomonas. These findings provide deep insights into the responding mechanism of anammox consortia to CIP stress and promote the application of anammox process for treating wastewater containing antibiotics.202438554504
790960.9992Simultaneous efficient removal of tetracycline and mitigation of antibiotic resistance genes enrichment by a modified activated sludge process with static magnetic field. To address the increasing issue of antibiotic wastewater, this study applied a static magnetic field (SMF) to the activated sludge process to increase the efficiency of tetracycline (TC) removal from swine wastewater and to reveal its enhanced mechanisms. The results demonstrated that the SMF-modified activated sludge process could achieve almost complete TC removal at sludge loading rates of 0.3 mg TC/g MLSS/d. Analysis of zeta potential and extracellular polymeric substances composition of the activated sludge revealed that SMF increased electrostatic interactions between TC and activated sludge and made activated sludge has much more binding sites, finally resulting in the increased TC biosorption. Metagenomic analysis showed that SMF promoted the enrichment of ammonia-oxidizing bacteria, TC-degrading bacteria, and aromatic compounds-degrading bacteria; it also enhanced ammonia monooxygenase- and cytochrome P450-mediated TC metabolism while upregulating functional genes associated with oxidase, reductase, and dehydrogenase - all contributing to increased TC biodegradation. Additionally, SMF mitigated the enrichment and spread of antibiotic resistance genes (ARGs) by decreasing the abundance of potential hosts of ARGs and inhibiting the upregulation of genes encoding ABC transporters and putative transposase. Based on these findings, this study demonstrates that magnetic field is an enhancement strategy with great potential to relieve the harmful impacts of the growing antibiotic wastewater problem on human health and the ecosystem.202439038424
791870.9992Robustness of the partial nitrification-anammox system exposing to triclosan wastewater: Stress relieved by extracellular polymeric substances and resistance genes. The partial nitrification-anammox (PN/A) process is a promising method for the treatment of municipal wastewater. It is necessary to clarify the responses of PN/A system to antimicrobial agent triclosan (TCS) widely existed in the influent of wastewater treatment plants. In this study, it was found that PN/A system was robust to cope with 0.5 mg/L TCS. Specifically, the control reactor reached 80% total nitrogen removal efficiency (TNRE) on day 107, while the reactor feeding with 0.5 mg/L TCS reached the same TNRE on day 84. The results of the activity test, high-throughput sequencing and DNA-based stable isotope probing showed that 0.5 mg/L TCS did not impede the performance of ammonia oxidizing archaea, ammonia oxidizing bacteria (Nitrosomonas) and anammox bacteria (Candidatus Brocadia and Ca. Kuenenia), but significant inhibited the nitrite oxidizing bacteria (Nitrospira and Ca. Nitrotoga) and denitrifying bacteria. The influent TCS led to the increase of EPS content and enrichment of four resistance genes (RGs) (intI1, sul1, mexB, and tnpA), which might be two principal mechanisms by which PN/A can resist TCS. In addition, functional bacteria carrying multiple RGs also contributed to the maintenance of PN/A system function. These findings improved the understandings of antimicrobial effects on the PN/A system.202234954146
797080.9992Environmental micro-molar H(2)O(2) reduces the efficiency of glyphosate biodegradation in soil. Glyphosate is one of the most widely used pesticides globally. The environmental micro-molar hydrogen peroxide (H(2)O(2))-driven Fenton reaction has been reported to degrade herbicides in natural water. However, the impact of micro-molar H(2)O(2) (50 μM) on the degradation of glyphosate in soil and glyphosate-degrading bacteria remains unclear. In this study, degradation of glyphosate in the sterilized and unsterilized soil system and MSM medium under micro-molar H(2)O(2) was investigated; bacterial diversity, enzyme activity and gene abundance in the soil following micro-molar H(2)O(2) addition were also investigated. The results indicated that the addition of micro-molar H(2)O(2) facilitated the degradation of glyphosate in a sterilized environment, resulting in a 76.30% decrease in glyphosate within 30 days. The degradation of glyphosate increased by 52.32% compared to the control treatment. However, in an unsterilized environment, the addition of micro-molar H(2)O(2) leads to a reduction in the biodegradation efficiency of glyphosate. Bacteria, enzymes and specific genes were found to be affected to varying degrees. Firstly, micro-molar H(2)O(2) affects the relative abundance of functional bacteria related to glyphosate degradation, such as Afipia, Microcoleus and Pseudomonas. Secondly, micro-molar H(2)O(2) resulted in a decrease in soil phosphatase activity. Thirdly, the expression of resistance genes was affected, particularly the glyphosate resistance gene aroA. The findings presented a novel research perspective on the degradation of soil glyphosate by micro-molar H(2)O(2).202439307340
785190.9992Breaking antibiotic resistance: Sunlight-powered calcium peroxide for dual bactericidal and genetic elimination. Antibiotic-resistant bacteria (ARB) and associated antibiotic resistance genes (ARGs) have emerged as critical waterborne contaminants, posing serious public health risks. This study proposes a disinfection strategy through sunlight powered calcium peroxide (CaO(2)) treatment that simultaneously inactivates ARB and degrades ARGs in aquatic environments. Solar irradiation combined with CaO(2) (3.0 mM) activates dual mechanisms: alkaline-driven microbial inactivation (pH increase from 6.4 to 8.2 within 30 min) and ROS-mediated oxidative damage (ROS: (•)OH, H(2)O(2), (1)O(2) and O(2)(•-)), achieving complete 5-log inactivation of tetracycline and sulfonamides-resistant E. coli (TSRE). ARGs (tetA and sul2) showed 70-80 % reduction in absolute abundance, although the log removal did not exceed 1-log. Compared to sunlight alone, the addition of CaO(2) significantly enhanced disinfection efficiency. Alkaline and ROS-induced oxidative stress caused membrane lipid breakdown, protein denaturation, and suppression of antioxidant enzymes, along with DNA damage, lipid peroxidation, and enzyme inactivation. These effects increased membrane permeability, impaired bacterial recovery by downregulating DNA repair genes, and disrupted cellular integrity, ultimately limiting ARGs persistence. These findings highlight the synergistic effect of alkaline and oxidative stress in effectively inactivating ARB and degrading ARGs, positioning sunlight powered CaO(2) as a promising, highly efficient disinfection strategy for environmental water treatment.202540876436
8540100.9991Metagenomic insights into the mechanism for the rapid enrichment and high stability of Candidatus Brocadia facilitated by Fe(Ⅲ). The rapid enrichment of anammox bacteria and its fragile resistance to adverse environment are the critical problems facing of anammox processes. As an abundant component in anammox bacteria, iron has been proved to promote the activity and growth of anammox bacteria in the mature anammox systems, but the functional and metabolic profiles in Fe(III) enhanced emerging anammox systems have not been evaluated. Results indicated that the relative abundance of functional genes involved in oxidative phosphorylation, nitrogen metabolism, cofactors synthesis, and extracellular polymers synthesis pathways was significantly promoted in the system added with 5 mg/L Fe(III) (R5). These enhanced pathways were crucial to energy generation, nitrogen removal, cell activity and proliferation, and microbial self-defense, thereby accelerating the enrichment of anammox bacteria Ca. Brocadia and facilitating their resistance to adverse environments. Microbial community analysis showed that the proportion of Ca. Brocadia in R5 also increased to 64.42 %. Hence, R5 could adapt rapidly to the increased nitrogen loading rate and increase the nitrogen removal rate by 108 % compared to the system without Fe(III) addition. However, the addition of 10 and 20 mg/L Fe(III) showed inhibitory effects on the growth and activity of anammox bacteria, which exhibited the lower relative abundance of Ca. Brocadia and unstable or even collapsed nitrogen removal performance. This study not only clarified the concentration range of Fe(III) that promoted and inhibited the enrichment of anammox bacteria, but also deepened our understanding of the functional and metabolic mechanisms underlying enhanced enrichment of anammox bacteria by Fe(III), providing a potential strategy to hasten the start-up of anammox from conventional activated sludge.202438309072
7911110.9991Biochar induced inhibitory effects on intracellular and extracellular antibiotic resistance genes in anaerobic digestion of swine manure. Distribution of intracellular (iARGs) and extracellular ARGs (eARGs) in manure anaerobic digestion (AD) process coupled with two types of biochar (BC and BP) were investigated. And the effects of biochar on the conjugation transfer of ARGs were explored by deciphering the interaction of biochar with bacterial stress responses, physiological metabolism and antibiotic resistances. Results showed that AD process could effectively remove all the detected eARGs with efficiency of 47.4-98.2%. The modified biochar (BP) with larger specific surface area (SSA) was propitious to decrease the absolute copy number of extracellular resistance genes. AD process could effectively remove iARGs by inhibiting the growth of host bacteria. The results of structural equation models (SEM) indicated that biochar put indirect influences on the fate of ARGs (λ = -0.23, P > 0.05). Analysis on oxidative stress levels, antioxidant capacity, DNA damage-induced response (SOS) response and energy generation process demonstrated that biochar induced the oxidative stress response of microorganisms and enhanced the antioxidant capacity of bacteria. The elevated antioxidant capacity negatively affected SOS response, amplified cell membrane damage and further weakened the energy generation process, resulted in the inhibition of horizontal transfer of ARGs.202235609652
7569120.9991Simultaneous elimination of amoxicillin and antibiotic resistance genes in activated sludge process: Contributions of easy-to-biodegrade food. Antibiotics are continuously released into aquatic environments and ecosystems where they accumulate, which increases risks from the transmission of antibiotic resistance genes (ARGs). However, it is difficult to completely remove antibiotics by conventional biological methods, and during such treatment, ARGs may spread via the activated sludge process. Easy-to-biodegrade food have been reported to improve the removal of toxic pollutants, and therefore, this study investigated whether such co-substrates may also decrease the abundance of ARGs and their transferal. This study investigated amoxicillin (AMO) degradation using 0-100 mg/L acetate sodium as co-substrate in a sequencing biological reactor. Proteobacteria, Bacteroidetes, and Actinobacteria were identified as dominant phyla for AMO removal and mineralization. Furthermore, acetate addition increased the abundances of adeF and mdsC as efflux resistance genes, which improved microbial resistance, the coping ability of AMO toxicity, and the repair of the damage from AMO. As a result, acetate addition contributed to almost 100% AMO removal and stabilized the chemical oxygen demand (~20 mg/L) in effluents when the influent AMO fluctuated from 20 to 100 mg/L. Moreover, the total abundance of ARGs decreased by approximately ~30%, and the proportion of the most dominant antibiotic resistance bacteria Proteobacteria decreased by ~9%. The total abundance of plasmids that encode ARGs decreased by as much as ~30%, implying that the ARG spreading risks were alleviated. In summary, easy-to-biodegrade food contributed to the simultaneous elimination of antibiotics and ARGs in an activated sludge process.202133757248
8527130.9991Insight into the impacts and mechanisms of ketone stress on the antibiotic resistance in Escherichia coli. Accumulation of toxic organic has posed a substantial pressure on the proliferation of bacterial resistance. While aromatic organics have been demonstrated to enhance the antibiotic resistance in bacteria, no information is yet available on the effects of non-aromatic organics on the variations of bacterial resistance. Here, we investigated the effects of a typical ketone (i.e., methylisobutanone (MIBK)) on the variations of antibiotic resistance in Escherichia coli (E. coli). The results showed that the growth of resistant E. coli under environmental concentration of 50 μg/L MIBK was firstly inhibited as explained by the transient disruption in the cell membrane and then recovered possibly due to the reactive oxygen species. Exposure to 50 μg/L MIBK gradually raised the abundance of representative resistance gene (ampR) in E. coli. In contrast, the high concentration of 50 mg/L MIBK continuously inhibited the growth of resistant E. coli by disrupting cell membrane and notably promoted the proliferation of ampR through enhancing the horizontal transformation and up-regulating the expression of efflux pump gene. These findings provided the first evidence for the evolution of bacterial resistance in response to ketone organics.202235771331
7908140.9991DNA-based stable isotope probing deciphered the active denitrifying bacteria and triclosan-degrading bacteria participating in granule-based partial denitrification process under triclosan pressure. Granule-based partial denitrification (PD) is a technology that can supply stable nitrite for applying anaerobic ammonia oxidation in wastewater treatment, and triclosan (TCS) is a frequently detected antibacterial agent in wastewater treatment plants, therefore it is possible that TCS could enter into wastewater that is treated using PD technology. However, the active microorganisms responsible for PD and TCS removing in granule-based PD system have not been clearly identified and it is currently not clear how TCS affects the PD process. In this study, the impacts of TCS on PD performance, PD microbial community, antibiotic resistance genes (ARGs), active PD bacteria and TCS-degrading bacteria in a granule-based PD system were investigated. 3 mg/L TCS had adverse influence on PD process, but PD system could recover gradually after inhibiting of 10 days. After a period of domestication, PD granular sludge could achieve 10.66% of TCS degradation efficiency and 43.62% of TCS adsorption efficiency. Microbes might increase their resistance to TCS by increasing the secretion of extracellular polymeric substances, and the secretion of protein might play a more pivotal role than the secretion of polysaccharides in resisting TCS. The short-term shock of TCS might cause the propagation of acrA-03, while the long-term operation of TCS could propagate fabK and intI1. DNA stable isotope probing assay indicated that Thauera was active PD bacteria and TCS-degrading bacteria in the granule-based PD system, and it could contribute to nitrite accumulation and TCS degradation, simultaneously.202234979468
8535150.9991Metagenomics combined with DNA-based stable isotope probing provide comprehensive insights of active triclosan-degrading bacteria in wastewater treatment. The biotransformation of triclosan (TCS) during wastewater treatment occurred frequently, while little researches are known the identity of microorganisms involved in the biodegradation process. In this work, DNA-based stable isotope probing (DNA-SIP) was occupied to investigate the TCS assimilation microbes originated from a full-scale cyclic activated sludge system in Beijing. Results of TCS removal pathway showed that the TCS removal in nitrification process was mainly contributed by the metabolism of heterotrophic bacteria, accounting for about 18.54%. DNA-SIP assay indicated that Sphingobium dominated the degradation of TCS. Oligotyping analysis further indicated that oligotype GCTAAT and ATGTTA of Sphingobium played important roles in degrading TCS. Furthermore, the Kyoto Encyclopedia of Genes and Genomes functional abundance statistics based on PICRUSt2 showed that glutathione transferase was the most prevalent enzyme involved in TCS metabolism, and TCS might be removed through microbial carbon metabolism. Metagenomics made clear that Sphingobium might play irrelevant role on the propagation of antibiotics resistance genes (ARGs), even though, it could degrade TCS. Thauera and Dechloromonas were identified as the key hosts of most ARGs. This study revealed the potential metabolic pathway and microbial ecology of TCS biodegradation in nitrification process of wastewater treatment system.202133069997
7981160.9991Dissolved biochar eliminates the effect of Cu(II) on the transfer of antibiotic resistance genes between bacteria. The proliferation of antibiotic resistance genes (ARGs) has posed significant risks to human and environmental health. Research has confirmed that Cu(II) could accelerate the conjugative transfer of ARGs between bacteria. This study found that adding dissolved biochar effectively weakened or eliminated the Cu(II)-facilitated efficient transfer of ARGs. The efficiency of conjugative transfer was promoted after treatment with Cu(II) (0.05 mg/L) or dissolved biochar at a pyrolysis temperature of 300 °C. When exposed to the combination of Cu(II) and dissolved biochar, the transfer frequency was significantly reduced; this occurred regardless of the Cu(II) concentration or pyrolysis temperature of dissolved biochar. In particular, when the Cu(II) concentration exceeded 0.5 mg/L, the transfer efficiency was entirely inhibited. Gene expression analysis indicated that different treatments affect transfer efficiency by regulating the expression of three global regulatory genes: korA, korB, and trbA. Among them, humic acid repressed the expression of these genes; however, Cu(II) formed complex with the humic acid-like components, gradually weakening the inhibitive effect of these components. The promotion of low molecule organic matters dominated, resulting in a dynamic decline in the transfer efficiency. This study provides a new environmental contaminant treatment approach to eliminate the heavy metal-facilitated transfer of ARGs between bacteria.202234583164
8507170.9991Co-effect of cadmium and iron oxide nanoparticles on plasmid-mediated conjugative transfer of antibiotic resistance genes. Conjunctive transfer of antibiotic resistance genes (ARGs) among bacteria driven by plasmids facilitated the evolution and spread of antibiotic resistance. Heavy metal exposure accelerated the plasmid-mediated conjunctive transfer of ARGs. Nanomaterials are well-known adsorbents for heavy metals removal, with the capability of combatting resistant bacteria/facilitating conjunctive transfer of ARGs. However, co-effect of heavy metals and nanomaterials on plasmid-mediated conjunctive transfer of ARGs was still unknown. In this study, we investigated the effect of the simultaneous exposure of Cd(2+) and nano Fe(2)O(3) on conjugative transfer of plasmid RP4 from Pseudomonas putida KT2442 to water microbial community. The permeability of bacterial cell membranes, antioxidant enzyme activities and conjugation gene expression were also investigated. The results suggested that the combination of Cd(2+) and high concentration nano Fe(2)O(3) (10 mg/L and 100 mg/L) significantly increased conjugative transfer frequencies of RP4 plasmid (p < 0.05). The most transconjugants were detected in the treatment of co-exposure to Cd(2+) and nano Fe(2)O(3), the majority of which were identified to be human pathogens. The mechanisms of the exacerbated conjugative transfer of ARGs were involved in the enhancement of cell membrane permeability, antioxidant enzyme activities, and mRNA expression levels of the conjugation genes by the co-effect of Cd(2+) and nano Fe(2)O(3). This study confirmed that the simultaneous exposure to Cd(2+)and nano Fe(2)O(3) exerted a synergetic co-effect on plasmid-mediated conjunctive transfer of ARGs, emphasizing that the co-effect of nanomaterials and heavy metals should be prudently evaluated when combating antibiotic resistance.202133798824
8506180.9991Extracellular Polymeric Substances Acting as a Permeable Barrier Hinder the Lateral Transfer of Antibiotic Resistance Genes. Antibiotic resistance genes (ARGs) in bacteria are emerging contaminants as their proliferation in the environment poses significant threats to human health. It is well recognized that extracellular polymeric substances (EPS) can protect microorganisms against stress or damage from exogenous contaminants. However, it is not clear whether EPS could affect the lateral transfer of ARGs into bacteria, which is one of the major processes for the dissemination of ARGs. This study investigated the lateral transfer of ARGs carried by plasmids (pUC19, pHSG298, and pHSG396) into competent Escherichia coli cells with and without EPS. Transformant numbers and transformation efficiency for E. coli without EPS were up to 29 times of those with EPS at pH 7.0 in an aqueous system. The EPS removal further increased cell permeability in addition to the enhanced cell permeability by Ca(2+), which could be responsible for the enhanced lateral transfer of ARGs. The fluorescence quenching experiments showed that EPS could strongly bind to plasmid DNA in the presence of Ca(2+) and the binding strength (LogK (A) = 10.65-15.80 L mol(-1)) between EPS and plasmids was positively correlated with the enhancement percentage of transformation efficiency resulting from the EPS removal. X-ray photoelectron spectroscopy (XPS) analyses and model computation further showed that Ca(2+) could electrostatically bind with EPS mainly through the carboxyl group, hydroxyl group, and RC-O-CR in glucoside, thus bridging the plasmid and EPS. As a result, the binding of plasmids with EPS hindered the lateral transfer of plasmid-borne ARGs. This study improved our understanding on the function of EPS in controlling the fate and transport of ARGs on the molecular and cellular scales.201931057498
7968190.9991Induced ciprofloxacin biotransformation and antibiotic-resistance genes control in sulfate-reducing microbial fuel cells: Strategy and mechanism. Ciprofloxacin-containing saline wastewater treatment gains increasing attentions, due to the problems of limited degradation and spreading risk of antibiotic-resistance genes (ARGs). Sulfate reduction is a cost-efficient technology for simultaneous sulfate and antibiotic removal. The microbial fuel cell enhances removal of antibiotics and reduces spreading risk of ARGs in effluents, however, the biotransformation of ciprofloxacin (CIP) in sulfate-reducing microbial fuel cell (SR-MFC) remains unclear. Thus, a SR-MFC is established in this study for treatment of CIP-containing saline wastewater, which achieves simultaneous removal of CIP (50.2%), sulfate (85.1%), and ARGs (17.0%). The Desulfovibrio sp. bacteria become dominant in free biomass (58.8%) and biofilm (73.6%) after CIP exposing, respectively. The CIP can be utilized in prior to lactate for sulfate reduction, while the energy production is initially contributed to sulfate reduction followed by sulfide oxidation. Notably, the expression of ARGs declines probably due to enhanced biotransformation and limited adsorption (2.6%) of CIP on biomass after CIP addition. Long-term exposure to CIP enriches the ARGs of antibiotic efflux pump, implying some CIP is pumped out from intracellular to extracellular. A novel degradation pathway attacking the N15 site in piperazine may be the major and environmental-friendly biotransformation reaction, where the enzyme of ammonia-lyase and acetyltransferase are involved in. To our best knowledge, this is the first report of the novel pathway in bacterial CIP degradation system, which is known as fungal CIP biotransformation pathway. This study provides insights for CIP biotransformation in SR-MFC, and the operational strategy for antibiotic-containing saline wastewater treatment with ARGs control.202540058044