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343400.9957Insights into microbial contamination and antibiotic resistome traits in pork wholesale market: An evaluation of the disinfection effect of sodium hypochlorite. Chlorine and its derivatives, such as sodium hypochlorite (NaClO) and chlorine dioxide, are frequently employed as disinfectants throughout the pork supply chain in China. Nevertheless, the extensive use of NaClO has the potential to cause the creation of 'chlorine-tolerant bacteria' and accelerate the evolution of antibiotic resistance. This study evaluated the efficacy of NaClO disinfection by examining alterations in the microbiome and resistome of a pork wholesale market (PWM), and bacteria isolation and analysis were performed to validate the findings. As expected, the taxonomic compositions of bacteria was significantly different before and after disinfection. Notably, Salmonella enterica (S. enterica), Salmonella bongori (S. bongori), Escherichia coli (E. coli), Klebsiella pneumoniae (K. pneumoniae), and Pseudomonas aeruginosa (P. aeruginosa) were observed on all surfaces, indicating that the application of NaClO disinfection treatment in PWM environments for pathogenic bacteria is limited. Correlations were identified between antibiotic resistance genes (ARGs) associated with aminoglycosides (aph(3'')-I, aph(6')-I), quinolone (qnrB, abaQ), polymyxin (arnA, mcr-4) and disinfectant resistance genes (emrA/BD, mdtA/B/C/E/F). Furthermore, correlations were found between risk Rank I ARGs associated with aminoglycoside (aph(3')-I), tetracycline (tetH), beta_lactam (TEM-171), and disinfectant resistance genes (mdtB/C/E/F, emrA, acrB, qacG). Importantly, we found that Acinetobacter and Salmonella were the main hosts of disinfectant resistance genes. The resistance mechanisms of the ARGs identified in PWM were dominated by antibiotic deactivation (38.7%), antibiotic efflux (27.2%), and antibiotic target protection (14.4%). The proportion of genes encoding efflux pumps in the PWM resistome increased after disinfection. Microbial cultures demonstrated that the traits of microbial contamination and antibiotic resistane were consistent with those observed by metagenomic sequencing. This study highlights the possibility of cross-resistance between NaClO disinfectants and antibiotics, which should not be ignored.202438382341
784310.9957Inactivation of chlorine-resistant bacteria (CRB) via various disinfection methods: Resistance mechanism and relation with carbon source metabolism. With the widespread use of chlorine disinfection, chlorine-resistant bacteria (CRB) in water treatment systems have gained public attention. Bacterial chlorine resistance has been found positively correlated with extracellular polymeric substance (EPS) secretion. In this study, we selected the most suitable CRB controlling method against eight bacterial strains with different chlorine resistance among chloramine, ozone, and ultraviolet (UV) disinfection, analyzed the resistance mechanisms, clarified the contribution of EPS to disinfection resistance, and explored the role of carbon source metabolism capacity. Among all the disinfectants, UV disinfection showed the highest disinfection capacity by achieving the highest average and median log inactivation rates for the tested strains. For Bacillus cereus CR19, the strain with the highest chlorine resistance, 40 mJ/cm(2) UV showed a 1.90 log inactivation, which was much higher than that of 2 mg-Cl(2)/L chlorine (0.67 log), 2 mg-Cl(2)/L chloramine (1.68 log), and 2 mg/L ozone (0.19 log). Meanwhile, the UV resistance of the bacteria did not correlate with EPS secretion. These characteristics render UV irradiation the best CRB controlling disinfection method. Chloramine was found to have a generally high inactivation efficiency for bacteria with high chlorine-resistance, but a low inactivation efficiency for low chlorine-resistant ones. Although EPS consumed up to 56.7% of chloramine which an intact bacterial cell consumed, EPS secretion could not explain chloramine resistance. Thus, chloramine is an acceptable CRB control method. Similar to chlorine, ozone generally selected high EPS-secreting bacteria, with EPS consuming up to 100% ozone. Therefore, ozone is not an appropriate method for controlling CRB with high EPS secretion. EPS played an important role in all types of disinfection resistance, and can be considered the main mechanism for bacterial chlorine and ozone disinfection resistance. However, as EPS was not the main resistance mechanism in UV and chloramine disinfection, CRB with high EPS secretion were inactivated more effectively. Furthermore, carbon source metabolism was found related to the multiple resistance of bacteria. Those with low carbon source metabolism capacity tended to have higher multiple resistance, especially to chlorine, ozone, and UV light. Distinctively, among the tested gram-negative bacteria, in contrast to other disinfectants, chloramine resistance was negatively correlated with EPS secretion and positively correlated with carbon source metabolism capacity, suggesting a special disinfection mechanism.202337659185
757020.9956Effect and mechanism of residual-chlorine on the horizontal transfer of antibiotic resistance genes of chlorine resistant bacteria in reclaimed water. Chlorine disinfection inactivates most microbes in reclaimed water, but chlorine resistant bacteria (CRB) persist, threatening water safety and spreading antibiotic resistance genes (ARGs). ARG proliferation in reclaimed water systems risks public health, as dissemination via irrigation or urban reuse may enable environmental transmission to humans, exacerbating global antibiotic resistance. One hundred and fifty-two strains of CRB were isolated from reclaimed water in this study, and the detection rate of ARGs in those CRB was 100%, the detection rate for blTEM was 100%, followed by sul3 and tetG. Macrogenomic analysis revealed that Proteobacteria, Actinobacteria, and Bacteroidetes are the dominant CRB in reclaimed water. Long-term induction with the minimum inhibitory concentration (MIC) of NaClO enhanced the resistance of CRB to both Amp and NaClO. The EPS of CRB increased 1.30- to 2.04-fold, and the elevated surface hydrophobicity may serve as a co-resistance mechanism. EPS hindered disinfectant/antibiotic penetration, while hydrophobicity reduced hydrophilic molecule adhesion and promoted bacterial aggregation, both of which contribute to the enhanced resistance of CRB. Residual chlorine dose-dependently enhances ARG conjugation via ROS-SOS, ATP, and EPS pathways, unveiling novel CRB mechanisms and urging revised disinfection to mitigate ARG spread.202540580511
719630.9956The drinking water treatment process as a potential source of affecting the bacterial antibiotic resistance. Two waterworks, with source water derived from the Huangpu or Yangtze River in Shanghai, were investigated, and the effluents were plate-screened for antibiotic-resistant bacteria (ARB) using five antibiotics: ampicillin (AMP), kanamycin (KAN), rifampicin (RFP), chloramphenicol (CM) and streptomycin (STR). The influence of water treatment procedures on the bacterial antibiotic resistance rate and the changes that bacteria underwent when exposed to the five antibiotics at concentration levels ranging from 1 to 100 μg/mL were studied. Multi-drug resistance was also analyzed using drug sensitivity tests. The results indicated that bacteria derived from water treatment plant effluent that used the Huangpu River rather than the Yangtze River as source water exhibited higher antibiotic resistance rates against AMP, STR, RFP and CM but lower antibiotic resistance rates against KAN. When the antibiotic concentration levels ranged from 1 to 10 μg/mL, the antibiotic resistance rates of the bacteria in the water increased as water treatment progressed. Biological activated carbon (BAC) filtration played a key role in increasing the antibiotic resistance rate of bacteria. Chloramine disinfection can enhance antibiotic resistance. Among the isolated ARB, 75% were resistant to multiple antibiotics. Ozone oxidation, BAC filtration and chloramine disinfection can greatly affect the relative abundance of bacteria in the community.201526150304
343040.9956Benzalkonium chloride alters phenotypic and genotypic antibiotic resistance profiles in a source water used for drinking water treatment. Antibiotic resistance is a major public health concern. Triclosan is an antimicrobial compound with direct links to antibiotic resistance that was widely used in soaps in the U.S. until its ban by the U.S. Food and Drug Administration. Benzalkonium chloride (BAC), a quaternary ammonium compound, has widely replaced triclosan in soaps marketed as an antibacterial. BAC has been detected in surface waters and its presence will likely increase following increased use in soap products. The objective of this study was to determine the effect of BAC on relative abundance of antibiotic resistance in a bacterial community from a surface water used as a source for drinking water treatment. Bench-scale microcosm experiments were conducted with microbial communities amended with BAC at concentrations ranging from 0.1 μg L(-1) to 500 μg L(-1). Phenotypic antibiotic resistance was quantified by culturing bacteria in the presence of different antibiotics, and genotypic resistance was determined using qPCR to quantify antibiotic resistance genes (ARGs). BAC at concentrations ranging from 0.1 μg L(-1) to 500 μg L(-1) was found to positively select for bacteria resistant to ciprofloxacin and sulfamethoxazole, and negatively select against bacteria with resistance to six other antibiotics. Exposure to BAC for 14 days increased the relative abundance of sul1 and bla(TEM). This study re-highlights the importance of employing both culture and non-culture-based techniques to identify selection for antibiotic resistance. The widespread use of BAC will likely impact antibiotic resistance profiles of bacteria in the environment, including in source waters used for drinking water, wastewater treatment plants, and natural waterways.202031767233
469850.9955Antidepressant exposure as a source of disinfectant resistance in waterborne bacteria. The emergence of disinfectant-resistant pathogens in water is a major threat to public health. However, whether human-consumed pharmaceuticals can induce bacterial resistance to disinfectants remains unclear. Herein, Escherichia coli was exposed to 12 antidepressants, and susceptibility of antidepressant-induced chloramphenicol (CHL)-resistant mutants to disinfectants was tested. Whole genome sequencing, global transcriptomic sequencing, and real-time quantitative polymerase chain reaction were used to elucidate the underlying mechanisms. We observed that duloxetine, fluoxetine, amitriptyline, and sertraline significantly increased the mutation frequency of E. coli against CHL by 15- to 2948-fold. The resultant mutants increased the average MIC(50) of sodium hypochlorite, benzalkonium bromide, and triclosan roughly 2- to 8-fold. Consistently, marRAB and acrAB-tolC genes, together with ABC transporter genes (e.g., yddA, yadG, yojI, and mdlA), were triggered to increase the efflux of disinfectants out of the cell, while ompF was inhibited, reducing disinfectant penetration into the cell. Additionally, the occurrence of DNA mutations in marR and acrR in the mutants was observed, potentially resulting in increased synthesis of the AcrAB-TolC pump. This study indicates that pharmaceutical exposure may create disinfectant-resistant bacteria, which may then be released into water systems, providing novel insights into the potential source of water-borne disinfectant-resistant pathogens.202337030229
850560.9955Environmental concentrations of surfactants as a trigger for climax of horizonal gene transfer of antibiotic resistance. Ubiquitous antibiotic resistance genes (ARGs) is a significant global human health concern. Surfactants have been extensively used worldwide, and the consumption of surfactants containing hygiene, cleaning agents and disinfectants was multiplied during COVID-19 pandemic, which have caused significantly increased pollution of surfactants in aquatic environment. Whether such ever-increasing surfactant concentration boost dissemination risk of ARGs still remains unknown. Here the effects of three typical surfactants such as sodium dodecyl sulfate, cetyltrimethylammonium bromide and benzalkonium chloride on the transformation of pUC19 plasmid (2686 bp)-borne ARGs to recipient bacteria E. coli DH5ɑ were investigated. It was found that these surfactants at environmental concentrations facilitated horizonal gene transfer (HGT) via transformation. The transformation triggering concentrations for the three surfactants were 0.25-0.34 mg/L with a maximum increased transformation frequency of 13.51-22.93-fold. The mechanisms involved in activated HGT of ARGs via transformation triggered by surfactants could be mainly attributed to the increased production of reactive oxygen species, which further enhanced cell membrane permeability. These findings provide new sights for understanding of ARG propagation and also imply that the drastic rise of surfactant concentration in aquatic environment may significantly increase the dissemination risk of antibiotic resistance.202337484423
787270.9954Quaternary ammonium compounds promoted anoxic sludge granulation and altered propagation risk of intracellular and extracellular antibiotic resistance genes. Surfactants could influence sludge morphology and disinfectants were linked to antibiotic resistance genes (ARGs). Thus, the response of activated sludge and ARGs to long-term quaternary ammonium compounds (QACs) exposure required further investigation, which is a popular surfactant and disinfectant. Here, three sequencing batch reactors were fed with 5 mg/L most frequently detected QACs (dodecyl trimethyl ammonium chloride (ATMAC C12), dodecyl benzyl dimethyl ammonium chloride (BAC C12) and didodecyl dimethyl ammonium chloride (DADMAC C12)) for 180 d. The long-term inhibitory effect on denitrification ranked: DADMAC C12 > BAC C12 > ATMAC C12. Besides, obvious granular sludge promoted by the increase of α-Helix/(β-Sheet + Random coil) appeared in DADMAC C12 system. Moreover, intracellular ARGs increased when denitrification systems encountered QACs acutely but decreased in systems chronically exposed to QACs. Although replication and repair metabolism in ATMAC C12 system was higher, ATMAC C12 significantly promoted proliferation of extracellular ARGs. It was noteworthy that the propagation risk of extracellular ARGs in sludge increased significantly during sludge granulation process, and intracellular sul2 genes in sludge and water both increased with the granular diameter in DADMAC C12 system. The universal utilization of QACs may enhance antibiotic resistance of bacteria in wastewater treatment plants, deserving more attention.202336444811
784680.9954Removal of antibiotic resistance genes and inactivation of antibiotic-resistant bacteria by oxidative treatments. The persistence of antibiotics in the environment because of human activities, such as seafood cultivation, has attracted great attention as they can give rise to antibiotic resistance genes (ARGs) and antibiotic-resistant bacteria (ARB). In this study, we explored the inactivation and removal efficiencies of Escherichia coli SR1 and sul1 (plasmid-encoded ARGs), respectively, in their extracellular and intracellular forms (eARGs and iARGs) by three commonly used fishery oxidants, namely chlorine, bromine, and potassium permanganate (KMnO(4)), at the practical effective concentration range (0.5, 5, and 15 mg/L). Kinetics data were obtained using laboratory phosphate-buffered saline (PBS). Following the same fishery oxidation methods, the determined kinetics models were tested by studying the SR1 and sul1 disinfection efficiencies in (sterilized) pond water matrix. At concentrations of 5 and 15 mg/L, all three oxidants achieved sufficient cumulative integrated exposure (CT values) to completely inactivate SR1 and efficiently remove sul1 (up to 4.0-log). The oxidation methods were then applied to an unsterilized pond water matrix in order to study and evaluate the indigenous ARB and ARGs disinfection efficiencies in aquaculture, which reached 1.4-log and 1.0-log during treatment with fishery oxidants used in pond preparation at high concentrations before stocking (5-15 mg/L), respectively. A high chlorine concentration (15 mg/L) could efficiently remove ARGs (or iARGs) from pond water, and the iARG removal efficiency was higher than that of eARGs in pond water. The method and results of this study could aid in guiding future research and practical disinfection to control the spread of ARGs and ARB in aquaculture.202134030387
361890.9954The role of the qacA gene in mediating resistance to quaternary ammonium compounds. Conditions facilitating resistance to quaternary ammonium compounds (QACs) were investigated in Staphylococcus aureus SK982 exposed to benzalkonium chloride (BAC; a member of QACs) under various circumstances. S. aureus SK982 carrying the qacA gene encoding for resistance to QACs was grown in the presence of stable or gradually increasing concentrations of BAC, or it was exposed to this antiseptic in the exponential phase of growth. Bacteria cultivated in the highest BAC concentrations that did not retard their growth comparing to the untreated control were subjected to real-time quantitative polymerase chain reaction analysis for relative expression of the efflux genes qacA and norA. Under such conditions, S. aureus SK982 tolerated a relatively low stable concentration of BAC (1.22 mg/L) when compared with a gradually increasing antiseptic concentration (tolerance of 4.88 mg/L). However, in both cases, qacA expression was not significant. The culture exposed in the exponential phase of growth tolerated the highest concentration of BAC (9.76 mg/L) as also accompanied by significant overexpression of qacA. Expression of norA was relatively low regardless of the conditions tested. It seems that under the short-term conditions, the phase of bacterial growth is more important for the expression of BAC resistance than the capability to adapt to this antiseptic. This study provides a deeper insight into the relevance of the qac genes in conferring resistance to QACs.201323256651
6751100.9954Assessment 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
6785110.9954Combined impact of antibiotics and Cr(VI) on antibiotic resistance, ARGs, and growth of Bacillussp. SH-1: A functionl analysis from gene to protease. The simultaneous selection of antibiotic resistance genes (ARGs) induced by heavy metals and antibiotics has emerged as a growing environmental problem. This study investigated the combined effects of chromium (Cr(VI)) and antibiotics on the ARGs of Bacillus cereus SH-1. As Cr(VI) concentration increased, it triggered reactive oxygen species oxidative stress in SH-1, increased antioxidant enzyme activity, enhanced plasmid conjugative transfer, and reduced the efficiency of Cr(VI) removal by SH-1. Antibiotic resistance varied with increasing tetracycline and amoxicillin minimum inhibitory concentrations (MICs), whereas azithromycin and chloramphenicol MICs decreased with Cr(VI) induction. The overexpression of eight genes of the HAE-1 family of efflux pumps was detected using metagenomics and proteomics. Co-contamination with Cr(VI) and antibiotics has led to the emergence and spread of antibiotic-resistant bacteria. Therefore, resistance gene contamination resulting from Cr(VI)-polluted environments cannot be overlooked.202439384050
6746120.9953Environmental concentration of the quaternary ammonium disinfectant benzalkonium chloride strongly induces resistance gene profiles in fish. Disinfectants are non-antibiotic biocides that have been used extensively in daily life, particularly since the onset of the COVID-19 pandemic. However, their effect on drug resistance has not received sufficient attention. Here, marine medaka were subjected to an environmental concentration (10 μg/L) of benzalkonium chloride (BAC), sulfamethazine (SMZ), and their combination, aiming to elucidate their contributions to antibiotic resistance. Overall, 10 μg/L BAC exhibited a stronger induction potential for multiple antibiotic resistance genes (ARGs) relative to a similar level of SMZ. Specifically, tetracycline resistance genes were readily induced, regardless of exposure to BAC, SMZ, or their combination. BAC exhibited a more pronounced induction of ARGs than SMZ and showed a stronger potential to stimulate multidrug resistance. SMZ and BAC induced distinct virulence factors. Bacteria increased pathogenicity primarily through biofilm formation and enhanced community sensing under SMZ exposure, whereas iron acquisition and the production of reactive oxygen species appeared to be the main mechanisms by which bacteria evaded host defenses under BAC exposure. A greater number of ARGs demonstrated a significant positive correlation with virulence factors following BAC exposure compared to both the SMZ exposure group and the co-exposure group, which further confirmed the strong ability of BAC to induce multidrug resistance. In summary, owing to the typically unregulated and low-dose use of disinfectants in daily life and their pseudo-persistence in the environment, their potential to induce resistance may exceed that of antibiotics. Therefore, increased attention and preventive measures are required to address their resistance-inducing effects.202540073566
7847130.9953Inactivation and change of tetracycline-resistant Escherichia coli in secondary effluent by visible light-driven photocatalytic process using Ag/AgBr/g-C(3)N(4). Control of antibiotic-resistant bacteria (ARB) and their related genes in secondary effluents has become a serious issue because of increased awareness of their health risks. A considerable number of techniques have been developed in recent years, particularly in relation to advanced oxidation. However, limited information is known about cellular behavior and resistance characteristic change during photocatalytic treatment. In this study, the inactivation of tetracycline (TC)-resistant Escherichia coli (TC-E. coli), removal of TC-resistant genes (TC-RGs), and antibiotic susceptibility were evaluated by employing photocatalytic treatment using Ag/AgBr/g-C(3)N(4) with visible light irradiation. The effects of light intensity, photocatalyst dosage, and reaction ambient temperature on photocatalysis were modelled and investigated. The rate of TC-E. coli removal was also optimized. Results demonstrated that the optimal conditions for TC-E. coli removal included light intensity of 96.0 mW/cm(2), photocatalyst dosage of 211.0 mg/L, and reaction ambient temperature of 23.7 °C. Under such conditions, the ARB removal rate was 6.1 log after 90 min and the related TC-RG removal rates were 49%, 86%, 69%, and 86% for tetA, tetM, tetQ, and intl1, respectively. The minimum inhibitory concentration test after photocatalysis shows that the antibiotic resistance of TC-E. coli was enhanced, which may be mainly due to the changes in the membrane potential and resulted in difficulty in destroying the bacteria through antibiotic contact. Hence, photocatalytic treatment could be an ideal method for ARB and antibiotic-resistant gene (ARG) control in wastewater, but the health risks of the remaining ARB and ARG should be investigated further.202031841919
8029140.9953Migration of antibiotic resistance genes and evolution of flora structure in the Xenopus tropicalis intestinal tract with combined exposure to roxithromycin and oxytetracycline. The intestinal flora is one of the most important environments for antibiotic resistance development, owing to its diverse mix of bacteria. An excellent medicine model organism, Xenopus tropicalis, was selected to investigate the spread of antibiotic resistance genes (ARGs) in the intestinal bacterial community with single or combined exposure to roxithromycin (ROX) and oxytetracycline (OTC). Seventeen resistance genes (tetA, tetB, tetE, tetM, tetO, tetS, tetX, ermF, msrA, mefA, ereA, ereB, mphA, mphB, intI1, intI2, intI3) were detected in the intestines of Xenopus tropicalis living in three testing tanks (ROX tanks, OTC tanks, ROX + OTC tanks) and a blank tank for 20 days. The results showed that the relative abundance of total ARGs increased obviously in the tank with single stress but decreased in the tank with combined stress, and the genes encoding the macrolide antibiotic efflux pump (msrA), phosphatase (mphB) and integron (intI2, intI3) were the most sensitive. With the aid of AFM scanning, DNA was found to be scattered short chain in the blank, became extended or curled and then compacted with the stress from a single antibiotic, and was compacted and then fragmented with combined stress, which might be the reason for the variation of the abundance of ARGs with stress. The ratio of Firmicutes/Bacteroides related to diseases was increased by ROX and OTC. The very significant correlation between intI2 and intI3 with tetS (p ≤ 0.001) hinted at a high risk of ARG transmission in the intestines. Collectively, our results suggested that the relative abundance of intestinal ARGs could be changed depending on the intestinal microbiome and DNA structures upon exposure to antibiotics at environmental concentrations.202235063519
7790150.9953Disinfection of polymicrobial urines by electrochemical oxidation: Removal of antibiotic-resistant bacteria and genes. In this work, data obtained from the University Hospital Complex of Albacete (Spain) were selected as a case study to carry out the disinfection experiments. To do this, different configurations of electrochemical reactors were tested for the disinfection of complex urines. Results showed that 4-6 logs bacterial removal were achieved for every bacterium tested when working with a microfluidic flow-through reactor after 180 min (0.423 Ah dm(-3)). The MIKROZON® cell reached a total disinfection after 60 min (1.212 Ah dm(-3)), causing severe damages induced in the cell walls observed in SEM images. The concentration profiles of the electrogenerated disinfectants in solution could explain the differences observed. Additionally, a mean decrease in the ARGs concentration ranked as follows: bla(KPC) (4.18-logs) > bla(TEM) (3.96-logs) > ermB (3.23-logs) using the MIKROZON® cell. This electro-ozonizer could be considered as a suitable alternative to reduce the risk of antibiotic resistance spread. Hence, this study provides an insight into different electrochemical reactors for the disinfection of complex hospital urine matrices and contributes to reduce the spread of antibiotic resistance through the elimination of ARGs. A topic of great importance nowadays that needs to be further studied.202234923384
3847160.9953The impact and mechanism of quaternary ammonium compounds on the transmission of antibiotic resistance genes. The emergence of antibiotic resistance genes (ARGs) in microbes can be largely attributed to the abuse and misuse of antibiotics and biocides. Quaternary ammonium compounds (QACs) have been used worldwide as common disinfectants and detergents; however, their potential impact on the spread and diffusion of ARGs is still unknown. In this study, we detected the QAC resistance gene (qacEΔ1), the 1 integron gene (intI1), and 12 ARGs (sul1, sul2, cfr, cml, fexA, tetA, tetG, tetQ, tetX, ermB, bla(TEM,) and dfrA1) in 48 water samples from three watersheds by quantitative PCR (qPCR). We investigated the evolution of bacterial antibiotic resistance under QAC and antibiotic environmental pressures by long-term continuous culture. In addition, five QACs were selected to investigate the effect of QAC on the efficiency of conjugation transfer. The changes in bacterial cell membrane and production of reactive oxygen species (ROS) were detected by flow cytometry, revealing the mechanism by which QAC affects the spread of antibiotic resistance. Our results showed that the QAC resistance gene was ubiquitous in watersheds and it had significant correlation with intI1 and seven ARGs (r = 0.999, p < 0.01). QACs could increase the resistance of bacteria to multiple antibiotics. Furthermore, all five QACs promoted the conjugation transfer of the RP4 plasmid; the optimal concentration of QACs was about 10(-1)-10(-2) mg/L and their transfer efficiencies were between 1.33 × 10(-6) and 8.87 × 10(-5). QACs enhanced membrane permeability of bacterial cells and stimulated bacteria to produce ROS, which potentially promoted the transfer of plasmids between bacteria. In conclusion, this study demonstrated that QACs may facilitate the evolution and gene transfer of antibiotic resistance gene among microbiome.201931372954
5752170.9953Cefoxitin inhibits the formation of biofilm involved in antimicrobial resistance MDR Escherichia coli. The study investigates the relationship between biofilm formation and antibiotic resistance in Escherichia coli (E. coli) isolated from calves. Using biochemical and molecular methods, we identified the isolates and assessed their biofilm-forming ability through an improved crystal violet staining method. The minimum inhibitory concentrations (MICs) of 18 antibiotics against the isolates were determined using the broth microdilution method. The impact of cefoxitin on biofilm formation was analyzed using laser scanning confocal microscopy (LSCM). Additionally, qRT-PCR was employed to evaluate the expression levels of biofilm-related genes (luxS, motA, fliA, pfs, and csgD) in response to varying cefoxitin concentrations. Results indicated a significant correlation between antimicrobial resistance (AMR) and biofilm formation ability. Cefoxitin effectively reduced biofilm formation of multidrug-resistant E. coli isolates at 1/2 and 1 MIC, with enhanced inhibition at higher concentrations. The QS-related genes luxS, pfs, motA, and fliA were downregulated, leading to decreased csgD expression. At 1/2 MIC, csgD expression was significantly reduced. In conclusion, cefoxitin inhibits biofilm formation in multidrug-resistant E. coli by down-regulating key genes, offering a potential strategy to mitigate resistance and control infections in calves caused by biofilm-positive E. coli isolates.202540122078
8065180.9953Synergistic enhancement effect of straw-earthworms in the reduction of sulfamethoxazole and antibiotic resistance genes. Soil antibiotic pollution is a global concern. It has been confirmed that straw or earthworm can enhance microbial degradation of antibiotics in soil. However, in the C/N transformation processes of soil ecosystems, straw and earthworms are closely interconnected. Whether their interaction can further enhance microbial degradation of antibiotic pollution and the underlying mechanisms remain to be explored. This study conducted a 90 days co-incubation experiment with four treatments: straw + earthworms + sulfamethoxazole (RS-EW-SMX), straw + SMX (RS-SMX), earthworms + SMX (EW-SMX), and SMX alone (SMX). Residual SMX, its degradation intermediates, and microbial communities were monitored at multiple timepoints. Results indicated an exponential decline in SMX degradation rates across treatments. By day 90, SMX was nearly completely degraded in all treatment groups. However, the combined effect of straw and earthworms significantly enhanced the degradation efficiency of SMX. During the rapid degradation phase, SMX in above four treatments decreased from 20.0 mg kg(-1) to 0.93, 1.88, 5.26 and 7.02 mg kg(-1), respectively at day 10. Furthermore, the RS-EW-SMX treatment promoted SMX transformation into low-molecular-weight intermediates and increased the relative abundance of SMX-degrading bacteria by 1.35, 2.01, and 2.17-fold compared to RS-SMX, EW-SMX, and SMX, respectively. SMX degradation efficiency exhibited a strong positive linear correlation with the relative abundance of degrading bacteria across all treatments (R(2) = 0.961). Concurrently, analysis revealed that straw presence facilitated the targeted enrichment of SMX-degrading bacteria within the earthworm gut, concomitant with a reduction in associated antibiotic resistance genes (ARGs). This synergistic interaction between straw and earthworms, mediated through the gut microbiome and carbon utilization, constitutes a primary mechanism underpinning the accelerated SMX degradation observed. These findings reveal a novel macrofauna-plant residues interaction mechanism for improved in situ antibiotic bioremediation, providing practical solutions for soil pollution mitigation.202540914087
7804190.9953Electrochemical flow-through disinfection reduces antibiotic resistance genes and horizontal transfer risk across bacterial species. Antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs), as emerging pollutants, are released into environment, increasing the risk of horizontal gene transfer (HGT). However, a limited number of studies quantified the effects of ARB disinfection on the HGT risk. This study investigated the inactivation of E. coli 10667 (sul) and the release and removal of ARGs using an electrochemical flow-through reactor (EFTR). Furthermore, the transfer frequencies and potential mechanisms of HGT after disinfection were explored using non-resistant E. coli GMCC 13373 as the recipient and E. coli DH5α carrying plasmid RP4 as the donor. A threshold of current density (0.25 mA/cm(2)) was observed to destroy cells and release intracellular ARGs (iARGs) to increase extracellular ARGs (eARGs) concentration. The further increase in the current density to 1 mA/cm(2) resulted in the decline of eARGs concentration due to the higher degradation rate of eARGs than the release rate of iARGs. The performance of ARGs degradation and HGT frequency by EFTR were compared with those of conventional disinfection processes, including chlorination and ultraviolet radiation (UV). A higher ARGs degradation (83.46%) was observed by EFTR compared with that under chlorination (10.23%) and UV (27.07%). Accordingly, EFTR reduced the HGT frequency (0.69) of released ARGs into the recipient (Forward transfer), and the value was lower than that by chlorination (2.69) and UV (1.73). Meanwhile, the surviving injured E. coli 10667 (sul) with increased cell permeability was transferred by plasmid RP4 from the donor (Reverse transfer) with a higher frequency of 0.33 by EFTR compared with that under chlorination (0.26) and UV (0.16). In addition, the sul3 gene was the least resistant to EFTR than sul1 and sul2 gene. These findings provide important insights into the mechanism of HGT between the injured E. coli 10667 (sul) and environmental bacteria. EFTR is a promising disinfection technology for preventing the spread of antibiotic resistance.202235085844