Metagenomics combined with DNA-based stable isotope probing provide comprehensive insights of active triclosan-degrading bacteria in wastewater treatment. - Related Documents




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853501.0000Metagenomics 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
853610.9996New insights into bioaugmented removal of sulfamethoxazole in sediment microcosms: degradation efficiency, ecological risk and microbial mechanisms. BACKGROUND: Bioaugmentation has the potential to enhance the ability of ecological technology to treat sulfonamide-containing wastewater, but the low viability of the exogenous degraders limits their practical application. Understanding the mechanism is important to enhance and optimize performance of the bioaugmentation, which requires a multifaceted analysis of the microbial communities. Here, DNA-stable isotope probing (DNA-SIP) and metagenomic analysis were conducted to decipher the bioaugmentation mechanisms in stabilization pond sediment microcosms inoculated with sulfamethoxazole (SMX)-degrading bacteria (Pseudomonas sp. M2 or Paenarthrobacter sp. R1). RESULTS: The bioaugmentation with both strains M2 and R1, especially strain R1, significantly improved the biodegradation rate of SMX, and its biodegradation capacity was sustainable within a certain cycle (subjected to three repeated SMX additions). The removal strategy using exogenous degrading bacteria also significantly abated the accumulation and transmission risk of antibiotic resistance genes (ARGs). Strain M2 inoculation significantly lowered bacterial diversity and altered the sediment bacterial community, while strain R1 inoculation had a slight effect on the bacterial community and was closely associated with indigenous microorganisms. Paenarthrobacter was identified as the primary SMX-assimilating bacteria in both bioaugmentation systems based on DNA-SIP analysis. Combining genomic information with pure culture evidence, strain R1 enhanced SMX removal by directly participating in SMX degradation, while strain M2 did it by both participating in SMX degradation and stimulating SMX-degrading activity of indigenous microorganisms (Paenarthrobacter) in the community. CONCLUSIONS: Our findings demonstrate that bioaugmentation using SMX-degrading bacteria was a feasible strategy for SMX clean-up in terms of the degradation efficiency of SMX, the risk of ARG transmission, as well as the impact on the bacterial community, and the advantage of bioaugmentation with Paenarthrobacter sp. R1 was also highlighted. Video Abstract.202438424602
790920.9996Simultaneous 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
790830.9996DNA-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
853440.9996Response of microbial nitrogen transformation processes to antibiotic stress in a drinking water reservoir. Effects of antibiotics on microbial nitrogen transformation processes in natural aquatic ecosystems are largely unknown. In this study, we utilized the (15)N stable isotope tracers and metagenomic sequencing to identify how antibiotics drive nitrogen transformation processes in Danjiangkou Reservoir, which is the largest artificial drinking water reservoir in China. We retrieved 51 nitrogen functional genes, and found that the highest abundances of nitrate reduction and denitrification-related genes occurred in dissimilatory nitrogen transformation pathways. (15)N-labelling analysis substantiated that denitrification was the main pathway for nitrogen removal, accounting for 57.1% of nitrogen loss. Nitrogen functional genes and antibiotic resistance genes co-occurred in Danjiangkou Reservoir, and they were mainly carried by the denitrifying bacteria such as Rhodoferax, Polaromonas, Limnohabitans, Pararheinheimera, Desulfobulbus, and Pseudopelobacter. Genome annotation revealed that antibiotic deactivation, Resistance-Nodulation-Division and facilitator superfamily efflux pumps were responsible for the multiple-resistance to antibiotics in these bacteria. Moreover, antibiotics showed non-significant effects on nitrogen transformation processes. It is speculated that denitrifying bacteria harboring ARGs played crucial roles in protecting nitrogen transformation from low-level antibiotics stress in the reservoir. Our results highlight that denitrifying bacteria are important hosts of ARGs, which provides a novel perspective for evaluating the effects of antibiotics on nitrogen cycle in natural aquatic ecosystems.202134303244
856650.9996Synergistic Control of Trimethoprim and the Antimicrobial Resistome in Electrogenic Microbial Communities. Synergistic control of the risks posed by emerging antimicrobials and antibiotic resistance genes (ARGs) is crucial for ensuring ecological safety. Although electrogenic respiration can enhance the biodegradation of several antimicrobials and reduce ARGs accumulation, the association mechanisms of antimicrobial biodegradation (trimethoprim, TMP) with the fate of the antimicrobial resistome remain unclear. Here, the biotransformation pathway of TMP, microbial associations, and functional gene profiles (e.g., degradation, antimicrobial resistance, and electron transfer) were analyzed. The results showed that the microbial electrogenic respiration significantly enhanced the biodegradation of TMP, especially with a cosubstrate sodium acetate supply. Electroactive bacteria enriched in the electrode biofilm positively correlated with potential TMP degraders dominated in the planktonic communities. These cross-niche microbial associations may contribute to the accelerated catabolism of TMP and extracellular electron transfer. Importantly, the evolution and dissemination of overall ARGs and mobile genetic elements (MGEs) were significantly weakened due to the enhanced cometabolic biodegradation of TMP. This study provides a promising strategy for the synergistic control of the water ecological risks of antimicrobials and their resistome, while also highlighting new insights into the association of antimicrobial biodegradation with the evolution of the resistome in an electrically integrated biological process.202438299532
856260.9996Nitrogen and phosphorus limitations promoted bacterial nitrate metabolism and propagation of antibiotic resistome in the phycosphere of Auxenochlorella pyrenoidosa. Despite that nitrogen (N) and phosphorus (P) play critical roles in the lifecycle of microalgae, how N and P further affect the distribution of bacteria and antibiotic resistance genes (ARGs) in the phycosphere is still poorly understood. In this study, the effects of N and P on the distribution of ARGs in the phycosphere of Auxenochlorella pyrenoidosa were investigated. Results showed that the growth and chlorophyll synthesis of microalgae were inhibited when N or P was limited, regardless of the N/P ratios, but the extracellular polymeric substances content and nitrate assimilation efficiency were enhanced in contrast. Metagenomic sequencing revealed that N or P limitation resulted in the recruitment of specific bacteria that highly contribute to the nitrate metabolism in the phycosphere. Besides, N or P limitation promoted the propagation of phycosphere ARGs, primarily through horizontal gene transfer mediated by mobile genetic elements. The enrichment of specific bacteria induced by changes in the algal physiology also contributed to the ARGs proliferation under nutrient limitation. Our results demonstrated that the reduction of algal cells caused by nutrient limitation could promote the propagation of ARGs, which provides new insights into the occurrence and spread of ARGs in the phycosphere.202438367442
856770.9996System-dependent divergence of microbial community and resistome in two anaerobic niches under sulfamethoxazole selection. The prevalence of sulfamethoxazole (SMX) in high-strength wastewater poses a significant threat to the stability and efficiency of anaerobic biological treatment systems, particularly when deployed as initial treatment units. However, the complex interactions arising from SMX biodegradation and their resultant effects on typical anaerobic digestion (AD) and sulfate-reducing (SR) systems are not thoroughly understood. This study revealed that SMX exposure stimulated methanogenesis in the AD system and sulfate reduction in the SR system, driven primarily by enriched key functional taxa (e.g., methanogens, sulfate-reducing bacteria). Organic matter removal efficiency increased significantly in the AD system under SMX stress, attributed to the enrichment of fermentative bacteria. Notably, the enriched class Actinomycetes was capable of SMX biodegradation, thereby likely mitigating SMX stress for other microorganisms. In contrast, the SR system exhibited significantly diminished organic matter removal despite developing a more functionally specialized community under SMX exposure. This community harbored fewer SMX degraders, perpetuating selective pressure on the microbiota. Increasing SMX concentrations failed to induce significant shifts in overall community structure in either system, while significantly promoted the proliferation of antibiotic resistance genes (ARGs), particularly pronounced in the SR system exhibiting high SMX accumulation. Moreover, mobile genetic elements mediated the horizontal transfer of the sulfonamide resistance gene sul1 and other co-occurring ARGs located on plasmids. This study provides novel insights into the convergent and divergent microbial responses in the AD and SR systems under SMX exposure, highlighting the dual effects (both stimulatory and inhibitory) of SMX on the functionality of these anaerobic systems.202541130171
673680.9995Biotic pathways of reciprocal responses between antibiotic resistance genes and inorganic nitrogen cycling genes in amoxicillin-stressed compost ecosystems. This study explored the transformation of inorganic nitrogen, the expression levels of antibiotic resistance genes (ARGs), and the regulatory mechanisms of key species on ARGs and inorganic nitrogen cycling genes (INCGs) under different levels of amoxicillin (AMX) stress. High level of AMX inhibited the accumulation of NH(4)(+)-N, which increased by 531 % relative to the initial. Moreover, AMX to some extent increased the levels of nirS and nirK, which could potentially result in nitrogen loss and the accumulation of NO(2)(-). Actinobacteria might serve as potential hosts for ARGs during sludge composting. This stress induced a complex response between INCGs and ARGs more complex due to key species. Under high-level AMX pressure, most species associated with ARGs likely derived from nitrogen cycling functional species. To conclude, high levels of AMX stress might lead to nitrogen cycling imbalance and the dissemination of antibiotic resistance genes in composting systems.202438387840
856390.9995Overlooked role of extracellular polymeric substances in antibiotic-resistance gene transfer within microalgae-bacteria system. Controlling the spread of antibiotic-resistance genes (ARGs) under antibiotic stress has become an increasingly urgent issue. Microalgae possess the capability to remove antibiotics while concurrently inhibiting ARGs. Microalgae-bacteria systems can produce significant quantities of extracellular polymeric substances (EPS). However, the roles of EPS in the spread of ARGs have not been sufficiently explored, resulting in an insufficient understanding of the contribution of each EPS component and a lack of analysis on the complex interactions between EPS and ARGs. This study systematically explored the overlooked role of EPS in the transmission of ARGs within microalgae-bacteria systems. The current results showed that the potential of the microalgae-bacteria system for treating antibiotic wastewater. The tightly bound-EPS (TB-EPS) can acquire the higher absolute abundances of ARGs compared with the loosely bound-EPS (LB-EPS). The correlation coefficient between polysaccharides and TB-EPS ARGs was higher than that between polysaccharides and LB-EPS ARGs. The gene patterns of LB-EPS closely clustered with those of TB-EPS, while intracellular ARG gene patterns differed from both TB-EPS and LB-EPS. Metagenomic analyses indicated that the relative abundances of sul1 and sul2 were considerably higher at the beginning stage compared to the end stage. The abundance of Achromobacter, increased by the end stage, aligning with its potential to produce exopolysaccharide. Additionally, the absolute abundance of genes encoding exopolysaccharides (nagB and galE) and conjugative transfer transcription regulator (traF), increased over time. These findings enhanced our comprehension of the significance of EPS on the fate of ARGs in microalgae-bacteria systems during the treatment of antibiotic-contaminated wastewater.202539879767
7907100.9995Determination of the fate of antibiotic resistance genes and the response mechanism of plants during enhanced antibiotic degradation in a bioelectrochemical-constructed wetland system. Chloramphenicol (CAP) has a high concentration and detection frequency in aquatic environments due to its insufficient degradation in traditional biological wastewater treatment processes. In this study, bioelectrochemical assistant-constructed wetland systems (BES-CWs) were developed as advanced processes for efficient CAP removal, in which the degradation and transfer of CAP and the fate of antibiotic resistance genes (ARGs) were evaluated. The CAP removal efficiency could reach as high as 90.2%, while the removed CAP can be partially adsorbed and bioaccumulated in plants, significantly affecting plant growth. The vertical gene transfer and horizontal gene transfer increased the abundance of ARGs under high voltage and CAP concentrations. Microbial community analysis showed that CAP pressure and electrical stimulation selected the functional bacteria to increase CAP removal and antibiotic resistance. CAP degradation species carrying ARGs could increase their opposition to the biotoxicity of CAP and maintain system performance. In addition, ARGs are transferred into the plant and upward, which can potentially enter the food chain. This study provides an essential reference for enhancing antibiotic degradation and offers fundamental support for the underlying mechanism and ARG proliferation during antibiotic biodegradation.202336931217
7958110.9995Microbial 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
7902120.9995Determination of the lower limits of antibiotic biodegradation and the fate of antibiotic resistant genes in activated sludge: Both nitrifying bacteria and heterotrophic bacteria matter. Antibiotics can be biodegraded in activated sludge via co-metabolism and metabolism. In this study, we investigated the biodegradation pathways of sulfamethoxazole (SMX) and antibiotic resistant genes' (ARGs) fate in different autotrophic and heterotrophic microorganisms, by employing aerobic sludge, mixed sludge, and nitrifying sludge. A threshold concentration of SMX activating the degradation pathways in the initial stage of antibiotics degradation was found and proved in different activated sludge systems. Heterotrophic bacteria played an important role in SMX biodegradation. However, ammonia-oxidizing bacteria (AOB) had a faster metabolic rate, which was about 15 times higher than heterotrophic bacteria, contributing much to SMX removal via co-metabolism. As SMX concentration increases, the amoA gene and AOB relative abundance decreased in aerobic sludge due to the enrichment of functional heterotrophic bacteria, while it increased in nitrifying sludge. Microbial community analysis showed that functional bacteria which possess the capacity of SMX removal and antibiotic resistance were selected by SMX pressure. Potential ARGs hosts could increase their resistance to the biotoxicity of SMX and maintain system performance. These findings are of practical significance to guide antibiotic biodegradation and ARGs control in wastewater treatment plants.202234799165
8541130.9995Insights into the response of anammox process to oxytetracycline: Impacts of static magnetic field. The long-term effects of oxytetracycline (OTC) with a high concentration on the anaerobic ammonium oxidation (Anammox) process were evaluated, and the role of static magnetic field (SMF) was further explored. The stress of OTC at 50 mg/L had little effect on the nitrogen removal of anammox process at the first 16 days. With the continuous addition of OTC and the increase of nitrogen loading, the OTC inhibited the nitrogen removal and anammox activity severely. During the 32 days of recovery period without OTC addition, the nitrogen removal was further deteriorated, indicating the inhibition of OTC on anammox activity was irreversible and persistent. The application of SMF alleviated the inhibition of OTC on anammox to some extent, and the specific anammox activity was enhanced by 47.1% compared to the system without SMF during the OTC stress stage. Antibiotic efflux was the major resistance mechanism in the anammox process, and tetA, tetG and rpsJ were the main functional antibiotic resistance genes. The addition of OTC weakened the metabolic interactions between the anammox bacteria and the symbiotic bacteria involved in the metabolism of cofactors and secondary metabolites, leading to the poor anammox activity. The adaptability of microbes to the OTC stress was improved by the application of SMF, which can enhance the metabolic pathways related to bacterial growth and resistance to environmental stress.202337586490
8540140.9995Metagenomic 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
7565150.9995Microalgae Enhances the Adaptability of Epiphytic Bacteria to Sulfamethoxazole Stress and Proliferation of Antibiotic Resistance Genes Mediated by Integron. The transmission of ARGs in the microalgae-associated epiphytic bacteria remains unclear under antibiotic exposure, apart from altering the microbial community structure. In this study, Chlorella vulgaris cocultured with bacteria screened from surface water was examined to explore the spread of ARGs in the presence of sulfamethoxazole (SMX). The extracellular polymers released by Chlorella vulgaris could reduce antibiotic-induced collateral damage to bacteria, thus increasing the diversity of the microalgae-associated epiphytic bacteria. The abundances of sul1 and intI1 in the phycosphere at 1 mg/L SMX dose increased by 290 and 28 times, respectively. Metagenomic sequencing further confirmed that SMX bioaccumulation stimulated the horizontal transfer of sul1 mediated by intI1 in the microalgae-associated epiphytic bacteria, while reactive oxygen species (ROS)-mediated oxidative stress induced the SOS response and thus enhanced the transformation of sul1 in the J group. This is the first study to verify that microalgae protect bacteria from antibiotic damage and hinder the spread of ARGs mediated by SOS response, while the transfer of ARGs mediated by integron is promoted due to the bioaccumulation of SMX in the phycosphere. The results contribute to present comprehensive understanding of the risk of ARG proliferation by the presence of emerging contaminants residues in river.202439417646
7566160.9995Antibiotic sulfadiazine degradation by persulfate oxidation: Intermediates dependence of ecotoxicity and the induction of antibiotic resistance genes. To preserve the water resources, this study has analyzed the ecotoxicity and antibiotic resistance genes (ARGs) induction capacity of sulfadiazine degradation intermediates resulting from persulfate activation oxidation enhanced by ultraviolet, ultrasound and microwave. The five degradation pathways caused by the contribution discrepancy of electron transfer and singlet oxygen ((1)O(2)) and variations in the ecotoxicity of different degradation products were analyzed. Microcosm experiment exhibited that the microbial community in actual water changed significantly with SDZ and degradation intermediates, in which the dominant genera were Aeromonas, Cupriavidus, Elizabethkingia and Achromobacter. Except for the selective pressure on bacteria, the degradation intermediates also exert a certain degree or even stronger induction on sulfonamide ARGs (sul4, sul1 and sul2) than SDZ. Furthermore, the potential hosts for sulfonamide ARGs were revealed by network analysis. These results provide a better understanding of antibiotics degradation mechanism and ARGs occurrence, which is useful for controlling the spread of ARGs.202336372382
7918170.9995Robustness 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
8565180.9995Deciphering the transfers of antibiotic resistance genes under antibiotic exposure conditions: Driven by functional modules and bacterial community. Antibiotics can exert selective pressures on sludge as well as affect the emergence and spread of antibiotic resistance genes (ARGs). However, the underlying mechanisms of ARGs transfers are still controversial and not fully understood in sludge system. In present study, two anaerobic sequence batch reactors (ASBR) were constructed to investigate the development of ARGs exposed to two sulfonamide antibiotics (SMs, sulfadiazine SDZ and sulfamethoxazole SMX) with increasing concentrations. The abundance of corresponding ARGs and total ARGs obviously increased with presence of SMs. Functional analyses indicated that oxidative stress response, signal transduction and type IV secretion systems were triggered by SMs, which would promote ARGs transfers. Network analysis revealed 18 genera were possible hosts of ARGs, and their abundances increased with SMs. Partial least-squares path modeling suggested functional modules directly influenced mobile genetic elements (MGEs) as well as the ARGs might be driven by both functional modules and bacteria community, while bacteria community composition played a more key role. Sludge with refractory antibiotics (SDZ) may stimulate the relevant functions and shift the microbial composition to a greater extent, causing more ARGs to emerge and spread. The mechanisms of ARGs transfers are revealed from the perspective of functional modules and bacterial community in sludge system for the first time, and it could provide beneficial directions, such as oxidative stress reduction, cellular communication control, bacterial composition directional regulation, for ARGs spread controlling in the future.202134563930
8564190.9995Effects of functional modules and bacterial clusters response on transmission performance of antibiotic resistance genes under antibiotic stress during anaerobic digestion of livestock wastewater. The formation and transmission of antibiotic resistance genes (ARGs) have attracted increasing attention. It is unclear whether the internal mechanisms by which antibiotics affect horizontal gene transfer (HGT) of ARGs during anaerobic digestion (AD) were influenced by dose and type. We investigated the effects of two major antibiotics (oxytetracycline, OTC, and sulfamethoxazole, SMX) on ARGs during AD according to antibiotic concentration in livestock wastewater influent. The low-dose antibiotic (0.5 mg/L) increased ROS and SOS responses, promoting the formation of ARGs. Meanwhile, low-dose antibiotics could also promote the spread of ARGs by promoting pili, communication responses, and the type IV secretion system (T4SS). However, different types and doses of antibiotics would lead to changes in the above functional modules and then affect the enrichment of ARGs. With the increasing dose of SMX, the advantages of pili and communication responses would gradually change. In the OTC system, low-dose has the strongest promoting ability in both pili and communication responses. Similarly, an increase in the dose of SMX would change T4SS from facilitation to inhibition, while OTC completely inhibits T4SS. Microbial and network analysis also revealed that low-dose antibiotics were more favorable for the growth of host bacteria.202336063716