Mechanism of earthworm coelomic fluid inhibits multidrug-resistant bacteria and blocks resistance transmission. - Related Documents




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678301.0000Mechanism of earthworm coelomic fluid inhibits multidrug-resistant bacteria and blocks resistance transmission. Antibiotic resistance is a growing global health crisis, especially the spread of multi-drug resistance. In this study, the inhibitory effects of earthworm coelomic fluid (ECF) on multidrug-resistant bacteria (MRB) were investigated during employing vermicomposting to treat excess sludge generated from wastewater treatment. The results demonstrated that the ECF was able to inhibit, even completely decompose the MRB. Notably, when the ECF concentration reached 1.0 mg/mL, the intracellular reactive oxygen species (ROS) level increased by 46.7 %, while cell viability decreased by 55.2 % compared to the control, demonstrating that ECF exerts strong antibacterial activity by inducing oxidative stress and disrupting cellular homeostasis. Furthermore, ECF effectively degraded the DNA of MRB, with removal rates of aphA, KanR, and tetA reaching 51.8 %, 42.3 %, and 35.0 %, respectively, indicating its ability to eliminate resistance genes and hinder their potential transfer. Additionally, the upregulation of genes involved in signaling, DNA replication and repair, and energy metabolism pathways suggests a systemic stress response in MRB, further supporting the broad-spectrum inhibitory effects of ECF on bacterial viability and resistance maintenance. Taken together, these findings may open a door to naturally and ecologically combat antibiotic resistance in pollutants control in wastewater treatment.202540706790
850110.9998Mechanistic insight of simultaneous removal of tetracycline and its related antibiotic resistance bacteria and genes by ferrate(VI). The emergence of antibiotics and their corresponding antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs) have posed great challenges to the public health. The paper demonstrates the removal of co-existing tetracycline (TC), its resistant Escherichia coli (E. coli), and ARGs (tetA and tetR) in a mixed system by applying ferrate(VI) (Fe(VI)O(4)(2-), Fe(VI)) at pH 7.0. TC was efficiently degraded by Fe(VI), and the rapid inactivation of the resistant E. coli was found with the complete loss of culturability. The results of flow cytometry suggested that the damage of membrane integrity and respiratory activity were highly correlated with the Fe(VI) dosages. Moreover, high-dose Fe(VI) eliminates 6 log(10) viable but non-culturable (VBNC) cells and even breaks the cells into fragments. ARGs in extracellular form (e-ARGs) exhibited a high sensitivity of 4.44 log(10) removal to Fe(VI). Comparatively, no removal of intracellular ARGs (i-ARGs) was observed due to the multi-protection of cellular structure and rapid decay of Fe(VI). The oxidized products of TC were assessed to be less toxic than the parent compound. Overall, this study demonstrated the superior efficiency and great promise of Fe(VI) on simultaneous removal of antibiotics and their related ARB and ARGs in water.202133984704
676120.9997Exposure to Al(2)O(3) nanoparticles facilitates conjugative transfer of antibiotic resistance genes from Escherichia coli to Streptomyces. The spread of antibiotic resistance genes (ARGs) has become a global environmental issue; it has been found that nanoparticles (NPs) can promote the transfer of ARGs between bacteria. However, it remains unclear whether NPs can affect this kind of conjugation in Streptomyces, which mainly conjugate with other bacteria via spores. In the present study, we demonstrated that Al(2)O(3) NPs significantly promote the conjugative transfer of ARGs from Escherichia coli (E. coli) ET12567 to Streptomyces coelicolor (S. coelicolor) M145 without the use of heat shock method. The number of transconjugants induced by Al(2)O(3) particles was associated with the size and concentration of Al(2)O(3) particles, exposure time, and the ratio of E. coli and spores. When nanoparticle size was 30 nm at a concentration of 10 mg/L, the conjugation efficiency reached a peak value of 182 cfu/10(8) spores, which was more than 60-fold higher than that of the control. Compared with nanomaterials, bulk particles exhibited no significant effect on conjugation efficiency. We also explored the mechanisms by which NPs promote conjugative transfer. After the addition of NPs, the intracellular ROS content increased and the expression of the classical porin gene ompC was stimulated. In addition, ROS enhanced the mRNA expression levels of conjugative genes by inhibiting global regulation genes. Meanwhile, expression of the conjugation-related gene intA was also stimulated, ultimately increasing the number of transconjugants. Our results indicated that Al(2)O(3) NPs significantly promoted the conjugative transfer of ARGs from bacteria to spores and aggravated the diffusion of resistance genes in the environment.201931561730
675130.9997Assessment of chlorine and hydrogen peroxide on airborne bacteria: Disinfection efficiency and induction of antibiotic resistance. Airborne pathogens severely threaten public health worldwide. Air disinfection is essential to ensure public health. However, excessive use of disinfectants may endanger environmental and ecological security due to the residual disinfectants and their by-products. This study systematically evaluated disinfection efficiency, induction of multidrug resistance, and the underlying mechanisms of disinfectants (NaClO and H(2)O(2)) on airborne bacteria. The results showed that airborne bacteria were effectively inactivated by atomized NaClO (>160 μg/L) and H(2)O(2) (>320 μg/L) after 15 min. However, some bacteria still survived after disinfection by atomized NaClO (0-80 μg/L) and H(2)O(2) (0-160 μg/L), and they exhibited significant increases in antibiotic resistance. The whole-genome sequencing of the resistant bacteria revealed distinct mutations that were responsible for both antibiotic resistance and virulence. This study also provided evidences and insights into possible mechanisms underlying the induction of antibiotic resistance by air disinfection, which involved intracellular reactive oxygen species formation, oxidative stress responses, alterations in bacterial membranes, activation of efflux pumps, and the thickening of biofilms. The present results also shed light on the role of air disinfection in inducing antibiotic resistance, which could be a crucial factor contributing to the global spread of antibiotic resistance through the air.202438823102
678140.9997Antibiotic-resistance gene transfer in antibiotic-resistance bacteria under different light irradiation: Implications from oxidative stress and gene expression. Due to the significant public health risks, there is substantial scientific interest in the increasing abundance of antibiotic-resistance bacteria (ARB) and the spread of antibiotic-resistance genes (ARGs) in aquatic environments. To clearly understand the mechanism of ARG transfer, this study examined the conjugative transfer of genes encoding resistance to cephalosporin (bla(CTX)) and polymyxin (mcr-1) from two antibiotic-resistant donor strains, namely E. coli DH5α (CTX) and E. coli DH5α (MCR), and to a streptomycin-resistant receptor strain (E. coli C600 (Sm)). Conjugative transfer was specifically studied under different light irradiation conditions including visible light (VL), simulated sunlight (SS) and ultraviolet light (UV(254nm)). Results show that the conjugative transfer frequency was not affected by VL irradiation, while it was slightly improved (2-10 fold) by SS irradiation and extremely accelerated (up to 100 fold) by UV irradiation. Furthermore, this study also explored the link between ARG transfer and stress conditions. This was done by studying physiological and biochemical changes; oxidative stress response; and functional gene expression of co-cultured AR-E. coli strains under stress conditions. When correlated with the transfer frequency results, we found that VL irradiation did not affect the physiological and biochemical characteristics of the bacteria, or induce oxidative stress and gene expression. For SS irradiation, oxidative stress occurred slowly, with a slight increase in the expression of target genes in the bacterial cells. In contrast, UV irradiation, rapidly inactivated the bacteria, the degree of oxidative stress was very severe and the expression of the target genes was markedly up-regulated. Our study could provide new insight into the underlying mechanisms and links between accelerated conjugative transfer and oxidative stress, as well as the altered expression of genes relevant to conjugation and other stress responses in bacterial cells.201930465986
674550.9997Decreased Antibiotic Susceptibility in Pseudomonas aeruginosa Surviving UV Irradition. Given its excellent performance against the pathogens, UV disinfection has been applied broadly in different fields. However, only limited studies have comprehensively investigated the response of bacteria surviving UV irradiation to the environmental antibiotic stress. Here, we investigated the antibiotic susceptibility of Pseudomonas aeruginosa suffering from the UV irradiation. Our results revealed that UV exposure may decrease the susceptibility to tetracycline, ciprofloxacin, and polymyxin B in the survival P. aeruginosa. Mechanistically, UV exposure causes oxidative stress in P. aeruginosa and consequently induces dysregulation of genes contributed to the related antibiotic resistance genes. These results revealed that the insufficient ultraviolet radiation dose may result in the decreased antibiotic susceptibility in the pathogens, thus posing potential threats to the environment and human health.202133613479
677560.9997Copper nanoparticles and copper ions promote horizontal transfer of plasmid-mediated multi-antibiotic resistance genes across bacterial genera. The spread of antibiotic resistance has become a major concern for public health. As emerging contaminants, various metallic nanoparticles (NPs) and ionic heavy metals have been ubiquitously detected in various environments. Although previous studies have indicated NPs and ionic heavy metals could exhibit co-selection effects for antibiotic resistance, little is known about whether and how they could promote antibiotic resistance spread via horizontal gene transfer across bacterial genera. This study, we report both CuO NPs and copper ions (Cu(2+)) could stimulate the conjugative transfer of multiple-drug resistance genes. When exposing bacteria to CuO NPs or Cu(2+) at environmental-relevant and sub-inhibitory concentrations (e.g., 1-100 μmol/L), conjugation frequencies of plasmid-encoded antibiotic resistance genes across genera (i.e., from Escherichia coli to Pseudomonas putida) were significantly enhanced (p < 0.05). The over-production of reactive oxygen species played a crucial role in promoting conjugative transfer. Genome-wide RNA and protein sequencing suggested expressional levels of genes and proteins related to oxidative stress, cell membrane permeability, and pilus generation were significantly up-regulated under CuO NPs and Cu(2+) exposure (p < 0.05). This study provides insights in the contributions of NPs and heavy metals on the spread of antibiotic resistance.201931158594
677370.9997Regulation of intracellular process by two-component systems: Exploring the mechanism of plasmid-mediated conjugative transfer. Plasmid-mediated conjugative transfer facilitates the dissemination of antibiotic resistance, yet the comprehensive regulatory mechanisms governing this process remain elusive. Herein, we established pure bacteria and activated sludge conjugation system to investigate the regulatory mechanisms of conjugative transfer, leveraging metformin as an exogenous agent. Transcriptomic analysis unveiled that substantial upregulation of genes associated with the two-component system (e.g., AcrB/AcrA, EnvZ/Omp, and CpxA/CpxR) upon exposure to metformin. Furthermore, downstream regulators of the two-component system, including reactive oxygen species (ROS), cytoplasmic membrane permeability, and adenosine triphosphate (ATP) production, were enhanced by 1.7, 1.4 and 1.1 times, respectively, compared to the control group under 0.1 mg/L metformin exposure. Moreover, flow sorting and high-throughput sequencing revealed increased microbial community diversity among transconjugants in activated sludge systems. Notably, the antibacterial potential of human pathogenic bacteria (e.g., Bacteroides, Escherichia-Shigella, and Lactobacillus) was augmented, posing a potential threat to human health. Our findings shed light on the spread of antibiotic resistance bacteria and assess the ecological risks associated with plasmid-mediated conjugative transfer in wastewater treatment systems.202438838482
762980.9997Graphene oxide in the water environment could affect tetracycline-antibiotic resistance. In recent years, the influence of new materials like nanoparticles in the water environment on biological substances has been widely studied. Antibiotic resistance genes (ARGs) represent a new type of pollutant in the environment. Graphene oxide (GO), as a nano material, because of its unique structure, may have an impact on antibiotic resistance bacteria (ARB) and ARGs; however the research in this area is rarely reported. Therefore, this study mainly investigated the effects of GO on bacterial antibiotic resistance. The results showed that GO had a limited effect on ARB inactivation. A high concentration of GO (>10 mg/L) can damage resistant plasmids to reduce bacterial resistance to antibiotics, but low concentrations of GO (<1 mg/L) led to almost no damage to the plasmid. However, all tested concentrations of GO promoted the conjugative transfer from 1to over 3 folds, with low concentrations and high concentration (1-10 and 100 mg/L) of GO samples the least promoted. The overall effect of GO on antibiotic resistance needs further investigation.201728549325
898190.9997Response mechanisms of different antibiotic-resistant bacteria with different resistance action targets to the stress from photocatalytic oxidation. The stress response of antibiotic-resistant bacteria (ARB) and the spread of antibiotic resistance genes (ARGs) pose a serious threat to the aquatic environment and human beings. This study mainly explored the effect of the heterogeneous photocatalytic oxidation (UVA-TiO(2) system) on the stress response mechanism of ARB with different antibiotic resistance action targets, including the cell wall, proteins, DNA, RNA, folate and the cell membrane. Results indicate that the stress response mechanism of tetracycline- and sulfamethoxazole-resistant E. coli DH5α, which targets the synthesis of protein and folate, could rapidly induce global regulators by the overexpression of relative antibiotic resistance action target genes. Different stress response systems were mediated via cross-protection mechanism, causing stronger tolerance to an adverse environment than other ARB. Moreover, the photocatalytic inactivation mechanism of bacterial cells and a graded response of cellular stress mechanism caused differences in the intensity of the stress mechanism of antibiotic resistance action targets. E. coli DH5α resistant to cefotaxime and polymyxin, targeting synthesis of the cell wall and cell membrane, respectively, could confer greater advantages to bacterial survival and higher conjugative transfer frequency than E. coli DH5α resistant to nalidixic acid and rifampicin, which target the synthesis of DNA and RNA, respectively. This new perspective provides detailed information on the practical application of photocatalytic oxidation for inactivating ARB and hampering the spreading of ARGs in the aquatic environment.202235453030
8502100.9997Simultaneously disinfection of amoebae, endosymbiotic bacteria, and resistance genes using a novel two-electron water oxidation strategy. Amoebae, which serve as important vectors for various pathogenic bacteria, are ubiquitous in natural and artificial water systems. Their robust survival capabilities and protective characteristics render conventional disinfection methods largely ineffective. Moreover, amoeba cells provide an ideal environment for the replication and transfer of antibiotic resistance genes, posing a significant threat to human health and safety. In this study, an in-situ activation system for electrocatalytic water oxidation was developed. This system effectively inactivates amoeba spores and their intracellular symbiotic bacteria while simultaneously reducing the abundance of resistance genes through the generation of hydroxyl radicals (•OH) and carbonate free radicals (•CO(3)(-)). The results demonstrated a 99.9 % inactivation rate for amoeba spores and a 99.999 % inactivation rate for intracellular bacteria. In addition, the prevalence of resistant genes in bacteria within amoebae, specifically including sul1 (sulfonamide resistance), tetA (tetracycline resistance), blaFOX (cefoxitin resistance), arsB (arsenic resistance), czcA (cadmium resistance), and copA (copper resistance), was significantly reduced by approximately 16 %-62.6 %. Therefore, this study introduces a new technology capable of simultaneously treating amoeba spores, intracellular bacteria, and resistance genes, which holds significant importance for reducing the spread of resistant genes and enhancing public health safety.202540449332
8516110.9997Graphene Oxide Inhibits Antibiotic Uptake and Antibiotic Resistance Gene Propagation. Antibiotics and antibiotic resistance genes (ARGs) in the natural environment have become substantial threats to the ecosystem and public health. Effective strategies to control antibiotics and ARG contaminations are emergent. A novel carbon nanomaterial, graphene oxide (GO), has attracted a substantial amount of attention in environmental fields. This study discovered the inhibition effects of GO on sulfamethoxazole (SMZ) uptake for bacteria and ARG transfer among microorganisms. GO promoted the penetration of SMZ from intracellular to extracellular environments by increasing the cell membrane permeability. In addition, the formation of a GO-SMZ complex reduced the uptake of SMZ in bacteria. Moreover, GO decreased the abundance of the sulI and intI genes by approximately 2-3 orders of magnitude, but the global bacterial activity was not obviously inhibited. A class I integron transfer experiment showed that the transfer frequency was up to 55-fold higher in the control than that of the GO-treated groups. Genetic methylation levels were not significant while sulI gene replication was inhibited. The biological properties of ARGs were altered due to the GO-ARG noncovalent combination, which was confirmed using multiple spectral analyses. This work suggests that GO can potentially be applied for controlling ARG contamination via inhibiting antibiotic uptake and ARG propagation.201627934199
8514120.9997Graphene oxide influences transfer of plasmid-mediated antibiotic resistance genes into plants. As an emerging contaminant, antibiotic resistance genes (ARGs) are raising concerns about its significant threat to public health. Meanwhile, graphene oxide (GO), which also has a potential ecological damage with increasingly entering the environment, has a great influence on the transfer of ARGs. However, little is known about the effects mechanisms of GO on the migration of antibiotic resistance genes (ARGs) from bacteria into plants. In this study, we investigated the influence of GO on the transfer of ARGs carried by RP4 plasmids from Bacillus subtilis into rice plants. Our results showed that the presence of GO at concentrations ranging from 0 to 400 mg L(-1) significantly reduced the transfer of ARGs into rice roots by 13-71 %. Moreover, the migration of RP4 from the roots to aboveground parts was significantly impaired by GO. These effects may be attributed to several factors. First, higher GO concentrations led to low pH in the culture solution, resulting in a substantial decrease in the number of antibiotic-resistant bacteria. Second, GO induced oxidative stress in rice, as indicated by enhanced Evans blue dye staining, and elevated levels of malondialdehyde, nitric oxide, and phenylalanine ammonia-lyase activity. The oxidative stress negatively affected plant growth, as demonstrated by the reduced fresh weight and altered lignin content in the rice. Microscopic observations confirmed the entry of GO into root cells but not leaf mesophyll cells. Furthermore, potential recipients of RP4 plasmid strains in rice after co-cultivation experiments were identified, including Bacillus subtilis, Bacillus amyloliquefaciens, and Bacillus cereus. These findings clarify the influence of GO on ARGs in the bacteria-plant system and emphasize the need to consider its potential ecological risks.202437979849
8819130.9997Responses of Bacillus sp. under Cu(II) stress in relation to extracellular polymeric substances and functional gene expression level. The production and composition of extracellular polymeric substances (EPS), as well as the EPS-related functional resistance genes and metabolic levels of Bacillus sp. under Cu(II) stress, were investigated. EPS production increased by 2.73 ± 0.29 times compared to the control when the strain was treated with 30 mg L(-1) Cu(II). Specifically, the polysaccharide (PS) content in EPS increased by 2.26 ± 0.28 g CDW(-1) and the PN/PS (protein/polysaccharide) ratio value increased by 3.18 ± 0.33 times under 30 mg L(-1) Cu(II) compared to the control. The increased EPS secretion and higher PN/PS ratio in EPS strengthened the cells' ability to resist the toxic effect of Cu(II). Differential expression of functional genes under Cu(II) stress was revealed by Gene Ontology pathway enrichment analysis and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis. The enriched genes were most obviously upregulated in the UMP biosynthesis pathway, the pyrimidine metabolism pathway, and the TCS metabolism pathway. This indicates an enhancement of EPS regulation-related metabolic levels and their role as a defense mechanism for cells to adapt to Cu(II) stress. Additionally, seven copper resistance genes were upregulated while three were downregulated. The upregulated genes were related to the heavy metal resistance, while downregulated genes were related to cell differentiation, indicating that the strain had initiated an obvious resistance to Cu(II) despite its severe cell toxicity. These results provided a basis for promoting EPS-regulated associated functional genes and the application of gene-regulated bacteria in heavy metal-containing wastewater treatment.202337195605
6748140.9997Time-dependent effects of ZnO nanoparticles on bacteria in an estuarine aquatic environment. Many studies have examined the acute toxicity of nanoparticles (NPs) towards model bacteria. In this study, we report the time-dependent effects of ZnO NPs on native, selected Zn-resistant and dominant bacteria in estuarine waters. An initial inhibition of bacterial growth followed by a recovery at 24 h was observed, and this rebound phenomenon was particularly notable when the raw water samples were treated with relatively high ZnO NP concentrations (1 and 10 mg/L).By comparing the groups treated with Zn(2+), Zn(2+) was shown to largely explain the acute cytotoxic effect of ZnO NPs on bacteria in raw waters. Furthermore, similar to the native bacteria, especially the dominant bacteria, the viability of Escherichia coli (E. coli) decreased with the increasing treatments time and the concentrations of ZnO NPs in water with different salinities. Moreover, the expression of Zn-resistance genes including zntA and zntR in E. coli suggested that the Zn-resistance system in E. coli can be activated to defend against the stress of Zn(2+) released from ZnO NPs, and salinity may promote this process in estuarine aquatic systems. Thus, the effect of ZnO NPs on bacteria in estuarine water bodies is likely determined by the synergistic effect of environmental salinity and dissolved Zn ions. As such, our findings are of high relevance and importance for understanding the ecological disturbances caused by anthropogenic NPs in estuarine environments.202031505343
8508150.9997Phenolic 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
7583160.9997Insights into the combined effect of ofloxacin and humic substances on sewage sludge anaerobic digestion. Humic substances (HS) and antibiotics are present simultaneously in various environments. However, the influence path and consequences of HS on antibiotics behaviors in complex anaerobic microbial systems are rarely known, hindering the understanding and control of antibiotics risks. This study for the first time investigated the combined effects of ofloxacin (OFL) and HS in sewage sludge anaerobic digestion system. Experimental results showed that OFL alone reduced the cumulative methane production and the maximum methane production rate by 14.6 % and 33.5 %, respectively. The methane production curves showed step by step adaption, which might be related with the increase of antibiotics resistance genes and their potential hosts. The coexistence of low concentration (6 % of sludge volatile solid) HS could alleviate the inhibition of OFL on hydrolysis-related bacteria and genes to a certain extent, thereby enhanced the methane production by 4.8 %. However, the coexistence of high concentration (12-24 % of sludge volatile solid) HS intensified the inhibition on hydrolysis-related bacteria and genes, and had more potential to combine with organic matters to prevent sludge solubilization, macromolecular organics hydrolysis and OFL degradation, thereby further decreasing the methane production by 7.6-15.9 %. Besides, the coexistence of OFL and high concentration HS increased the antimicrobial resistance and pathogenicity risks of digested sludge, by enhancing the residual level of verified pathogens, antibiotics resistance genes and virulence factor genes. This study provides new insights into the environmental risks of combined antibiotics and HS pollution, and offers a basis for strengthening the safe treatment and disposal of sewage sludge.202540752562
7581170.9997Enhanced performance of anaerobic digestion of cephalosporin C fermentation residues by gamma irradiation-induced pretreatment. Antibiotic fermentation residues is a hazardous waste due to the existence of residual antibiotics and antibiotic resistance genes (ARGs), probably leading to the induction and spread of antibiotic resistant bacteria (ARB) in the environment, which could pose potential harm to the ecosystem and human health. It is urgent to develop an effective technology to remove the residual antibiotics and ARGs. In this study, the anaerobic digestion combined with gamma irradiation was applied for the disposal and utilization of cephalosporin C fermentation residues. The experimental results showed that the antibacterial activities of cephalosporin C against Staphylococcus aureus were significantly decreased after anaerobic digestion. The removal of tolC, a multidrug resistant gene, was improved up to 100% by the combination of gamma irradiation and anaerobic digestion compared to solely anaerobic digestion process, which may be due to the changes of microbial community structures induced by gamma irradiation.202031590081
6750180.9997Viable but non-culturable E. coli induced by low level chlorination have higher persistence to antibiotics than their culturable counterparts. Disinfectant used in drinking water treatment and distribution system can induce culturable bacteria, including various kinds of pathogenic bacteria, into viable but non-culturable (VBNC) state. The loss of cultural state, resuscitation and environmental persistence of VBNC bacteria will severely damage drinking water microbiological safety and thus pose a risk to public health. The manner in which chlorination treatment induced a VBNC state in Escherichia coli and the antibiotic persistence of VBNC bacteria was investigated. It was found that low dosage of chlorine (0.5 mg L(-1)) disinfection effectively reduced the culturability of E. coli and induced a VBNC state, after which metabolic activity was reduced and persistence to 9 typical antibiotics was enhanced. Furthermore, RT-qPCR results showed that stress resistance genes (rpoS, marA, ygfA, relE) and ARGs, especially efflux genes were up-regulated compared with culturable cells. The intracellular concentration was tested and found to be lower in VBNC cells than in actively growing E. coli, which suggested a higher efflux rate. The data presented indicate that VBNC E. coli are more persistent than culturable counterparts to a wide variety of antibiotics. VBNC E. coli constitute a potential source of contamination and should be considered during monitoring of drinking water networks.201728662489
7958190.9997Microbial 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