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50400.8474Activation of Dithiolopyrrolone Antibiotics by Cellular Reductants. Dithiolopyrrolone (DTP) natural products are broad-spectrum antimicrobial and anticancer prodrugs. The DTP structure contains a unique bicyclic ene-disulfide that once reduced in the cell, chelates metal ions and disrupts metal homeostasis. In this work we investigate the intracellular activation of the DTPs and their resistance mechanisms in bacteria. We show that the prototypical DTP holomycin is reduced by several bacterial reductases and small-molecule thiols in vitro. To understand how bacteria develop resistance to the DTPs, we generate Staphylococcus aureus mutants that exhibit increased resistance to the hybrid DTP antibiotic thiomarinol. From these mutants we identify loss-of-function mutations in redox genes that are involved in DTP activation. This work advances the understanding of how DTPs are activated and informs development of bioreductive disulfide prodrugs.202539665630
50710.8440Tellurite resistance and reduction by obligately aerobic photosynthetic bacteria. Seven species of obligately aerobic photosynthetic bacteria of the genera Erythromicrobium, Erythrobacter, and Roseococcus demonstrated high-level resistance to tellurite and accumulation of metallic tellurium crystals. High-level resistance without tellurite reduction was observed for Roseococcus thiosulfatophilus and Erythromicrobium ezovicum grown with certain organic carbon sources, implying that tellurite reduction is not essential to confer tellurite resistance.199616535446
812720.8411Microbial Multitrophic Communities Drive the Variation of Antibiotic Resistome in the Gut of Soil Woodlice (Crustacea: Isopoda). Multitrophic communities inhabit in soil faunal gut, including bacteria, fungi, and protists, which have been considered a hidden reservoir for antibiotic resistance genes (ARGs). However, there is a dearth of research focusing on the relationships between ARGs and multitrophic communities in the gut of soil faunas. Here, we studied the contribution of multitrophic communities to variations of ARGs in the soil woodlouse gut. The results revealed diverse and abundant ARGs in the woodlouse gut. Network analysis further exhibited strong connections between key ecological module members and ARGs, suggesting that multitrophic communities in the keystone ecological cluster may play a pivotal role in the variation of ARGs in the woodlouse gut. Moreover, long-term application of sewage sludge significantly altered the woodlice gut resistome and interkingdom communities. The variation portioning analysis indicated that the fungal community has a greater contribution to variations of ARGs than bacterial and protistan communities in the woodlice gut after long-term application of sewage sludge. Together, our results showed that changes in gut microbiota associated with agricultural practices (e.g., sewage sludge application) can largely alter the gut interkingdom network in ecologically relevant soil animals, with implications for antibiotic resistance, which advances our understanding of the microecological drivers of ARGs in terrestrial ecosystem.202235876241
811730.8407Composting of oxytetracycline fermentation residue in combination with hydrothermal pretreatment for reducing antibiotic resistance genes enrichment. Hydrothermal pretreatment can efficiently remove the residual antibiotics in oxytetracycline fermentation residue (OFR), but its effect on antibiotic resistance genes (ARGs) during composting remains unclear. This study compared the shifts in bacterial community and evolutions in ARGs and integrons during different composting processes of OFRs with and without hydrothermal pretreatment. The results demonstrated that hydrothermal pretreatment increased the bacterial alpha diversity at the initial phase, and increased the relative abundances of Proteobacteria and Actinobacteria but decreased that of Bacteroidetes at the final phase by inactivating mycelia and removing residual oxytetracycline. Composting process inevitably elevated the abundance and relative abundance of ARGs. However, the increase in ARGs was significantly reduced by hydrothermal pretreatment, because the removal of oxytetracycline decreased their potential host bacteria and inhibited their horizontal gene transfer. The results demonstrated that hydrothermal pretreatment is an efficient strategy to reduce the enrichment of ARGs during the OFR composting.202033099099
849640.8407Neglected resistance risks: Cooperative resistance of antibiotic resistant bacteria influenced by primary soil components. Various antibiotic resistant bacteria (ARB) can thrive in soil and resist such environmental pressures as antibiotics through cooperative resistance, thereby promoting ARB retention and antibiotic resistance genes transmission. However, there has been finite knowledge in regard to the mechanisms and potential ecological risks of cooperative resistance in soil microbiome. In this study, soil minerals and organic matters were designed to treat a mixture of two Escherichia coli strains with different antibiotic resistance (E. coli DH5α/pUC19 and E. coli XL2-Blue) to determine how soil components affected cooperative resistance, and Luria-Bertani plates containing two antibiotics were used to observe dual-drug resistant bacteria (DRB) developed via cooperative resistance. Results showed quartz, humic acid, and biochar promoted E. coli XL2-Blue with high fitness costs, whereas kaolin, montmorillonite, and soot inhibited both strains. Using fluorescence microscope and PCR, it was speculated DRB could resist the antibiotic pressure via E. coli XL2-Blue coating E. coli DH5α/pUC19. E. coli DH5α/pUC19 dominated cooperative resistance. Correlation analysis and scanning electron microscope images indicated soil components influenced cooperative resistance. Biochar promoted cooperative resistance by increasing intracellular reactive oxygen species, thereby reducing the dominant strain concentration required for DRB development. Kaolin inhibited cooperative resistance the most, followed by soot and montmorillonite.202235074748
50550.8406Production of phytoalexins in peanut (Arachis hypogaea) seed elicited by selected microorganisms. Under favorable conditions, the peanut plant demonstrates appreciable resistance to fungal invasion by producing and accumulating phytoalexins, antimicrobial stilbenoids. This mechanism for resistance is little understood, yet it is crucial for breeding and genetically modifying peanut plants to develop new cultivars with fungal resistance. The dynamics of phytoalexin production in peanut seeds and embryos challenged by selected important fungi and bacteria was investigated. Different biotic agents selectively elicited production of major peanut stilbenoids, resveratrol, arachidin-1, arachidin-3, and SB-1. Aspergillis species, compared to other biotic agents, were more potent elicitors of stilbenoids. Embryos demonstrated significantly higher production of stilbenoids compared to cotyledons and may serve as a convenient source of genetic material in isolating genes for peanut plant defense enhancement.201323387286
812660.8402Antiallergic drugs drive the alteration of microbial community and antibiotic resistome in surface waters: A metagenomic perspective. Antiallergic drugs (AADs) are emerging contaminants of global concern due to their environmental persistence and potential ecological impacts. This study investigated the effects of seven AADs (chlorpheniramine, diphenhydramine, cetirizine, loratadine, desloratadine, sodium cromoglicate and calcium gluconate) at environmentally relevant concentrations on antibiotic resistome and bacterial community structures in water using microcosm experiments and metagenomic sequencing. The results showed that AADs increased the abundance of antibiotic-resistant bacteria (ARB) by 1.24- to 7.78-fold. Community structure shifts indicated that chlorpheniramine, diphenhydramine, and cetirizine promoted Actinobacteria (e.g., Aurantimicrobium), while the other four AADs favored Proteobacteria (e.g., Limnohabitans). AADs also significantly altered the relative abundance of antibiotic resistance genes (ARGs), with Actinobacteria and Proteobacteria identified as key ARB components and potential hosts of ARGs (e.g., evgS, mtrA, RanA). Host analysis showed ARGs were primarily carried by Actinobacteria (e.g., Aurantimicrobium) under chlorpheniramine, diphenhydramine, and cetirizine exposure, but by Proteobacteria (e.g., Limnohabitans) under the other four AADs. Furthermore, AADs facilitated the horizontal transfer of ARGs (e.g., evgS) within microbial communities, contributing to antibiotic resistance dissemination. This study highlights the ecological risks of AADs in promoting antibiotic resistance spread and provides new insights into their impact on microbial communities and resistome dynamics in aquatic environments.202540570627
811670.8401Effect of homemade compound microbial inoculum on the reduction of terramycin and antibiotic resistance genes in terramycin mycelial dreg aerobic composting and its mechanism. In order to tackle the issue of terramycin mycelial dreg (TMD) diagnosis and removal of terramycin and antibiotic resistance genes (ARGs), this study adopted aerobic composting (AC) technology and added homemade compound microbial inoculum (HCMI) to promote the AC of TMD and enhance the removal of terramycin and ARGs. The findings demonstrated that terramycin residue could be basically harmless after AC. Moreover, HCMI not only reduced QacB and tetH but also increased the degradation rates of VanRA, VanT, and dfrA24 by 40.81%, 5.65%, and 54.18%, respectively. The HCMI improved the removal rate of ARG subtypes to a certain extent. According to redundancy analysis, during AC, the succession of the microbial community had a stronger influence on the variance of ARG subtype than the environmental conditions. Differences in the abundance of various bacteria due to changes in temperature may be an intrinsic mechanism for the variation of ARG subtypes.202336403916
792680.8396Microplastics Exacerbated Conjugative Transfer of Antibiotic Resistance Genes during Ultraviolet Disinfection: Highlighting Difference between Conventional and Biodegradable Ones. Microplastics (MPs) have been confirmed as a hotspot for antibiotic resistance genes (ARGs) in wastewater. However, the impact of MPs on the transfer of ARGs in wastewater treatment remains unclear. This study investigated the roles and mechanisms of conventional (polystyrene, PS) and biodegradable (polylactic acid, PLA) MPs in the conjugative transfer of ARGs during ultraviolet disinfection. The results showed that MPs significantly facilitated the conjugative transfer of ARGs compared with individual ultraviolet disinfection, and PSMPs exhibited higher facilitation than PLAMPs. The facilitation effects were attributed to light shielding and the production of reactive oxygen species (ROS) and nanoplastics from ultraviolet irradiation of MPs. The light shielding of MPs protected the bacteria and ARGs from ultraviolet inactivation. More importantly, ROS and nanoplastics generated from irradiated MPs induced intracellular oxidative stress on bacteria and further increased the cell membrane permeability and intercellular contact, ultimately enhancing the ARG exchange. The greater fragmentation of PSMPs than PLAMPs resulted in a higher intracellular oxidative stress and a stronger enhancement. This study highlights the concerns of conventional and biodegradable MPs associated with the transfer of ARGs during wastewater treatment, which provides new insights into the combined risks of MPs and ARGs in the environment.202539723446
849790.8394Conjugation-mediated transfer of antibiotic resistance genes influenced by primary soil components and underlying mechanisms. Soil is the main natural reservoir of antibiotic resistant bacteria and antibiotic resistance genes (ARGs). Their dissemination and proliferation were largely motivated by conjugative transfer, while the influence of soil components on bacterial conjugative transfer and the underlying mechanisms remain poorly understood. In the present study, two Escherichia coli strains were exposed to soil minerals (quartz, kaolinite and montmorillonite) and organic matters (humic acid, biochar and soot) respectively to investigate their impact on ARGs conjugation. The results showed that quartz had no significant effect on conjugation; montmorillonite promoted the growth of the donor, but inhibited the recipient and conjugant; kaolinite and three organic matters significantly promoted the production of conjugant, while biochar promoted and then inhibited it with time prolong. Within the range of bacterial concentration involved in this study, the concentration of conjugant increased with the ratio of the concentration of donor and recipient (R(D/R)), indicating that the variation of conjugant production was mainly mediated by changing R(D/R). Further observation of biochar treatment group showed that the bacterial responses such as cell membrane permeability, cell surface hydrophobicity and biofilm formation ability shifted with the exposure time, which might be a potential factor affecting conjugative transfer. Collectively, our findings suggest that the type and exposure time of soil components jointly affected conjugation, while the change of R(D/R) and related bacterial responses are the main underlying mechanisms.202336586689
8119100.8394Biochar-amended composting of lincomycin fermentation dregs promoted microbial metabolism and reduced antibiotic resistance genes. Improper disposal of antibiotic fermentation dregs poses a risk of releasing antibiotics and antibiotic resistant bacteria to the environment. Therefore, this study evaluated the effects of biochar addition to lincomycin fermentation dregs (LFDs) composting. Biochar increased compost temperature and enhanced organic matter decomposition and residual antibiotics removal. Moreover, a 1.5- to 17.0-fold reduction in antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs) was observed. Adding biochar also reduced the abundances of persistent ARGs hosts (e.g., Streptomyces, Pseudomonas) and ARG-related metabolic pathways and genes (e.g., ATP-binding cassette type-2 transport, signal transduction and multidrug efflux pump genes). By contrast, compost decomposition improved due to enhanced metabolism of carbohydrates and amino acids. Overall, adding biochar into LFDs compost reduced the proliferation of ARGs and enhanced microbial community metabolism. These results demonstrate that adding biochar to LFDs compost is a simple and efficient way to decrease risks associated with LFDs composting.202336334868
329110.8392Effect of NlpE overproduction on multidrug resistance in Escherichia coli. NlpE, an outer membrane lipoprotein, functions during envelope stress responses in Gram-negative bacteria. In this study, we report that overproduction of NlpE increases multidrug and copper resistance through activation of the genes encoding the AcrD and MdtABC multidrug efflux pumps in Escherichia coli.201020211889
8619120.8391Bioavailability of pollutants and chemotaxis. The exposure of bacteria to pollutants induces frequently chemoattraction or chemorepellent reactions. Recent research suggests that the capacity to degrade a toxic compound has co-evolved in some bacteria with the capacity to chemotactically react to it. There is an increasing amount of data which show that chemoattraction to biodegradable pollutants increases their bioavailability which translates into an enhancement of the biodegradation rate. Pollutant chemoreceptors so far identified are encoded on degradation or resistance plasmids. Genetic engineering of bacteria, such as the transfer of chemoreceptor genes, offers thus the possibility to optimize biodegradation processes.201322981870
7887130.8390Double-edged sword effects of sulfate reduction process in sulfur autotrophic denitrification system: Accelerating nitrogen removal and promoting antibiotic resistance genes spread. This study proposed the double-edged sword effects of sulfate reduction process on nitrogen removal and antibiotic resistance genes (ARGs) transmission in sulfur autotrophic denitrification system. Excitation-emission matrix-parallel factor analysis identified the protein-like fraction in soluble microbial products as main endogenous organic matter driving the sulfate reduction process. The resultant sulfide tended to serve as bacterial modulators, augmenting electron transfer processes and mitigating oxidative stress, thereby enhancing sulfur oxidizing bacteria (SOB) activity, rather than extra electron donors. The cooperation between SOB and heterotroph (sulfate reducing bacteria (SRB) and heterotrophic denitrification bacteria (HDB)) were responsible for advanced nitrogen removal, facilitated by multiple metabolic pathways including denitrification, sulfur oxidation, and sulfate reduction. However, SRB and HDB were potential ARGs hosts and assimilatory sulfate reduction pathway positively contributed to ARGs spread. Overall, the sulfate reduction process in sulfur autotrophic denitrification system boosted nitrogen removal process, but also increased the risk of ARGs transmission.202439122125
7973140.8389Microplastic and antibiotic proliferated the colonization of specific bacteria and antibiotic resistance genes in the phycosphere of Chlorella pyrenoidosa. Despite that the phycosphere was an important niche for the proliferation of various bacteria and antibiotic resistance genes (ARGs), the factors that affect the colonization of bacteria and ARGs in the phycosphere are still poorly understood. In this study, sterile C. pyrenoidosa co-cultured with bacteria from different sources and provided with polylactic acid microplastic (PLA MPs) and florfenicol (FF) was examined. Results showed that bacteria promoted the growth of C. pyrenoidosa and increased its chlorophyll contents. PLA MPs and FF also showed positive effects on C. pyrenoidosa due to the "Hormesis effect". The occurrence of bacteria in the phycosphere was significantly affected by their sources and the addition of PLA MPs and FF. However, the core microbiota of the phycosphere in each group was similar. Additionally, PLA MPs and FF proliferated the abundance of phenicol-related ARGs (especially floR) and mobile genetic elements in the phycosphere. Notably, PLA MPs and FF enhanced the abundance of Flavobacterium, a potential host of ARGs. Our results highlighted the important roles of bacteria in microalgae and demonstrated exogenous pollutants could promote the spread of ARGs between surrounding environments and the phycosphere, which provide new insights into the occurrence and spread of ARGs in the phycosphere.202337201280
8488150.8388Antihistamine drug loratadine at environmentally relevant concentrations promotes conjugative transfer of antibiotic resistance genes: Coeffect of oxidative stress and ion transport. Due to the widespread use of loratadine (LOR) as an antihistamine, it is widely distributed in the environment as an emerging contaminant. However, its impact on the dissemination of antibiotic resistance genes (ARGs) remains unclear. This study investigated the effect of LOR on the conjugative transfer of ARGs and elucidated the potential mechanisms through transcriptome analysis. The results showed that LOR significantly promoted the frequency of conjugative transfer up to 1.5- to 8.6-fold higher compared with the control group. Exposure to LOR increased reactive oxidative species (ROS) and intracellular Ca(2+) concentrations, leading to the upregulation of expression of genes related to transmembrane transport and SOS response. Meanwhile, it stimulated the increase of cell membrane permeability. Moreover, LOR exposure could enhance H(+) efflux in donor bacteria, resulting in the decrease of intracellular pH and the elevation of transmembrane potential, which could induce the increase of ion transport, thereby promoting plasmid efflux from the cell membrane. Based on this, we inferred that LOR can induce an increase in ROS level and intracellular Ca(2+) concentrations, and promoted the efflux of intracellular H(+). This, in turn, triggered the intensification of various ion transport processes on the cell membrane, thereby increasing membrane permeability and accelerating plasmid efflux. Ultimately, the coeffect of oxidative stress response and ion transport promoted conjugative transfer. This study demonstrated that LOR significantly promotes plasmid-mediated conjugative transfer of ARGs, providing novel insights into the mechanisms underlying this process.202539919578
6938160.8386Assessment of the Effects of Biodegradable and Nonbiodegradable Microplastics Combined with Pesticides on the Soil Microbiota. Microplastics (MPs) and pesticides pose significant threats to the health of soil ecosystems. This study investigated the individual and combined effects of biodegradable polylactic acid (PLA) and nonbiodegradable polyethylene terephthalate (PET) microplastics alongside glyphosate and imidacloprid pesticides on soil microbial communities and antibiotic resistance genes (ARGs) via microcosm experiments. Compared with the control, PLA significantly increased microbial alpha diversity and enhanced microbial functions related to environmental information processing and metabolism. However, PLA also selectively enriched populations of beneficial and potentially pathogenic bacteria, whereas PET had comparatively weaker effects. Crucially, PLA exposure resulted in substantially higher total abundance and ecological risk levels of soil ARGs than did PET. Coexposure with pesticides further amplified these effects, with PLA demonstrating notable synergistic interactions with both glyphosate and imidacloprid. These findings challenge the conventional assumption that biodegradable MPs such as PLA are environmentally safer than nonbiodegradable MPs, thus highlighting their potential to induce more complex and potentially severe ecological risks under co-contamination scenarios with pesticides.202541175058
502170.8385A highly specialized flavin mononucleotide riboswitch responds differently to similar ligands and confers roseoflavin resistance to Streptomyces davawensis. Streptomyces davawensis is the only organism known to synthesize the antibiotic roseoflavin, a riboflavin (vitamin B2) analog. Roseoflavin is converted to roseoflavin mononucleotide (RoFMN) and roseoflavin adenine dinucleotide in the cytoplasm of target cells. (Ribo-)Flavin mononucleotide (FMN) riboswitches are genetic elements, which in many bacteria control genes responsible for the biosynthesis and transport of riboflavin. Streptomyces davawensis is roseoflavin resistant, and the closely related bacterium Streptomyces coelicolor is roseoflavin sensitive. The two bacteria served as models to investigate roseoflavin resistance of S. davawensis and to analyze the mode of action of roseoflavin in S. coelicolor. Our experiments demonstrate that the ribB FMN riboswitch of S. davawensis (in contrast to the corresponding riboswitch of S. coelicolor) is able to discriminate between the two very similar flavins FMN and RoFMN and shows opposite responses to the latter ligands.201222740651
8824180.8385Lactic acid bacteria modulate the CncC pathway to enhance resistance to β-cypermethrin in the oriental fruit fly. The gut microbiota of insects has been shown to regulate host detoxification enzymes. However, the potential regulatory mechanisms involved remain unknown. Here, we report that gut bacteria increase insecticide resistance by activating the cap "n" collar isoform-C (CncC) pathway through enzymatically generated reactive oxygen species (ROS) in Bactrocera dorsalis. We demonstrated that Enterococcus casseliflavus and Lactococcus lactis, two lactic acid-producing bacteria, increase the resistance of B. dorsalis to β-cypermethrin by regulating cytochrome P450 (P450) enzymes and α-glutathione S-transferase (GST) activities. These gut symbionts also induced the expression of CncC and muscle aponeurosis fibromatosis. BdCncC knockdown led to a decrease in resistance caused by gut bacteria. Ingestion of the ROS scavenger vitamin C in resistant strain affected the expression of BdCncC/BdKeap1/BdMafK, resulting in reduced P450 and GST activity. Furthermore, feeding with E. casseliflavus or L. lactis showed that BdNOX5 increased ROS production, and BdNOX5 knockdown affected the expression of the BdCncC/BdMafK pathway and detoxification genes. Moreover, lactic acid feeding activated the ROS-associated regulation of P450 and GST activity. Collectively, our findings indicate that symbiotic gut bacteria modulate intestinal detoxification pathways by affecting physiological biochemistry, thus providing new insights into the involvement of insect gut microbes in the development of insecticide resistance.202438618721
7645190.8383Antibiotic Resistance in the Collembolan Gut Microbiome Accelerated by the Nonantibiotic Drug Carbamazepine. The effects of pharmaceuticals as emerging contaminants in soil on the gut microbiome and antibiotic resistome in nontarget soil fauna are largely elusive. In this study, we explored the composition of the bacterial community and the presence of antibiotic resistance genes (ARGs) in the gut of the model soil collembolan (Folsomia candida) upon antiepileptic drug carbamazepine (CBZ) and antibiotic tetracycline (TC) exposure. Results showed that, individually or in combination, exposure to TC or CBZ significantly altered the gut community structure of F. candida, causing some enrichment of the bacteria associated with xenobiotic metabolism, such as Arthrobacter, Achromobacter, Gordonia, and Shinella. More importantly, oral exposure to the nonantibiotic drug CBZ enhanced the abundance and diversity of ARGs in the gut of F. candida, especially for the beta-lactams and multidrug resistance genes. Our results revealed that the most likely hosts of ARGs in the gut of F. candida were Proteobacteria and Actinobacteria. The significant positive correlation between mobile genetic elements (MGEs) and ARGs indicated the potential risk of ARGs transmission in the gut of F. candida. Overall, the nonantibiotic CBZ is likely to disturb the gut microbiota of nontarget soil fauna such as collembolans, thereby enhancing the dissemination of ARGs.202032816468