Metallic nanoparticles and ions accelerate the uptake of extracellular antibiotic resistance genes through transformation

2020 ◽  
Author(s):  
Jianhua Guo ◽  
Shuai Zhang ◽  
Ji Lu ◽  
Yue Wang ◽  
Willy Verstraete ◽  
...  
Keyword(s):  
Heavy Metal ◽  
Antibiotic Resistance ◽  
Cell Membrane ◽  
Resistance Genes ◽  
Natural Transformation ◽  
Antibiotic Resistance Genes ◽  
Acinetobacter Baylyi ◽  
Ag Nps ◽  
Zno Nps ◽  
Cuo Nps

Abstract Background: Antibiotic resistance genes (ARGs), heavy metal ions and nanoparticles (NPs) are emerging and ubiquitous contaminants in the environment. However, little is known about whether heavy metal-based NPs or ions could facilitate the dissemination of ARGs through natural transformation. This study evaluated the contributions of heavy metal-based NPs (Ag NPs, CuO NPs and ZnO NPs) and their ion forms (Ag + , Cu 2+ and Zn 2+ ) to the transformation of extracellular ARGs in Acinetobacter baylyi ADP1. Results: We found that these commonly-used NPs and ions from environmentally relevant concentrations can significantly promote the natural transformation frequency of ARGs by a factor of 11.0-folds, which is comparable to the effects of antibiotics. The enhanced transformation by Ag NPs, CuO NPs, Ag + and Cu 2+ was primarily associated with reactive oxygen species (ROS) over-production and cell membrane damage, which was also evident from up-regulations of both transcription and translation of ROS and outer membrane-related genes. Additionally, transmission electron microscope imaging revealed the roughened cell membrane after Ag NPs, CuO NPs, Ag + and Cu 2+ exposure. ZnO NPs and Zn 2+ might increase the natural transformation rate by stimulating the stress response and ATP synthesis. All tested NPs and ions resulted in up-regulating the competence and SOS response-associated genes. Conclusions: Our results demonstrate that Ag, CuO and ZnO-based NPs/ions from environmental concentrations could promote the natural transformation of plasmid-encoded ARGs into naturally competent A. baylyi . Our findings provide insights into the contributions of heavy metals and NPs to the spread of antibiotic resistance.

scholarly journals Chlorine disinfection facilitates natural transformation through ROS-mediated oxidative stress

2021 ◽  
Author(s):  
Shuai Zhang ◽  
Yue Wang ◽  
Ji Lu ◽  
Zhigang Yu ◽  
Hailiang Song ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Natural Transformation ◽  
Membrane Damage ◽  
Antibiotic Resistance Genes ◽  
Free Chlorine ◽  
Water Disinfection ◽  
Resistant Bacteria ◽  
Residual Chlorine ◽  
Acinetobacter Baylyi

AbstractThe bacterial infection that involves antimicrobial resistance is a rising global threat to public health. Chlorine-based water disinfection processes can inactivate antibiotic resistant bacteria. However, at the same time, these processes may cause the release of antibiotic resistance genes into the water as free DNA, and consequently increase the risk to disseminate antibiotic resistance via natural transformation. Presently, little is known about the contribution of residual chlorine affecting the transformation of extracellular antibiotic resistance genes (ARGs). This study investigates whether chloramine and free chlorine promote the transformation of ARGs and how this may occur. We reveal that both chloramine and free chlorine, at practically relevant concentrations, significantly stimulated the transformation of plasmid-encoded ARGs by the recipient Acinetobacter baylyi ADP1, by up to a 10-fold increase. The underlying mechanisms underpinning the increased transformations were revealed. Disinfectant exposure induced a series of cell responses, including increased levels of reactive oxygen species (ROS), bacterial membrane damage, ROS-mediated DNA damage, and increased stress response. These effects thus culminated in the enhanced transformation of ARGs. This promoted transformation was observed when exposing disinfectant-pretreated A. baylyi to free plasmid. In contrast, after pretreating free plasmid with disinfectants, the transformation of ARGs decreased due to the damage of plasmid integrity. These findings provide important insight on the roles of disinfectants affecting the horizontal transfer of ARGs, which could be crucial in the management of antibiotic resistance in our water systems.

scholarly journals Artificial sweeteners stimulate horizontal transfer of extracellular antibiotic resistance genes through natural transformation

2021 ◽  
Author(s):  
Zhigang Yu ◽  
Yue Wang ◽  
Ian R. Henderson ◽  
Jianhua Guo
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Horizontal Transfer ◽  
Natural Transformation ◽  
Model Organism ◽  
Antibiotic Resistance Genes ◽  
Artificial Sweeteners ◽  
Dna Uptake ◽  
Long Term Effects ◽  
Acinetobacter Baylyi

AbstractAntimicrobial resistance has emerged as a global threat to human health. Natural transformation is an important pathway for horizontal gene transfer, which facilitates the dissemination of antibiotic resistance genes (ARGs) among bacteria. Although it is suspected that artificial sweeteners could exert antimicrobial effects, little is known whether artificial sweeteners would also affect horizontal transfer of ARGs via transformation. Here we demonstrate that four commonly used artificial sweeteners (saccharin, sucralose, aspartame, and acesulfame potassium) promote transfer of ARGs via natural transformation in Acinetobacter baylyi ADP1, a model organism for studying competence and transformation. Such phenomenon was also found in a Gram-positive human pathogen Bacillus subtilis and mice faecal microbiome. We reveal that exposure to these sweeteners increases cell envelope permeability and results in an upregulation of genes encoding DNA uptake and translocation (Com) machinery. In addition, we find that artificial sweeteners induce an increase in plasmid persistence in transformants. We propose a mathematical model established to predict the long-term effects on transformation dynamics under exposure to these sweeteners. Collectively, our findings offer insights into natural transformation promoted by artificial sweeteners and highlight the need to evaluate these environmental contaminants for their antibiotic-like side effects.

scholarly journals Non-antibiotic pharmaceuticals can enhance the spread of antibiotic resistance via conjugation

2019 ◽  
Author(s):  
Yue Wang ◽  
Ji Lu ◽  
Shuai Zhang ◽  
Jie Li ◽  
Likai Mao ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Gene Transfer ◽  
Cell Membrane ◽  
Membrane Permeability ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
Protein Sequencing ◽  
Cell Membrane Permeability ◽  
Lipid Lowering ◽  
Potential Contribution

AbstractAntibiotic resistance is a global threat for public health. It is widely acknowledged that antibiotics at sub-inhibitory concentrations are important in disseminating antibiotic resistance via horizontal gene transfer. While there is high use of non-antibiotic human-targeted pharmaceuticals in our societies, the potential contribution of these on the spread of antibiotic resistance has been overlooked so far. Here, we report that commonly consumed non-antibiotic pharmaceuticals, including nonsteroidal anti-inflammatories (ibuprofen, naproxen, diclofenac), a lipid-lowering drug (gemfibrozil), and a β-blocker (propanolol), at clinically and environmentally relevant concentrations, significantly accelerated the conjugation of plasmid-borne antibiotic resistance genes. We looked at the response to these drugs by the bacteria involved in the gene transfer through various analyses that included monitoring reactive oxygen species (ROS) and cell membrane permeability by flow cytometry, cell arrangement, and whole-genome RNA and protein sequencing. We found the enhanced conjugation correlated well with increased production of ROS and cell membrane permeability. We also detected closer cell-to-cell contact and upregulated conjugal genes. Additionally, these non-antibiotic pharmaceuticals caused the bacteria to have responses similar to those detected when exposed to antibiotics, such as inducing the SOS response, and enhancing efflux pumps. The findings advance our understanding of the bacterial transfer of antibiotic resistance genes, and importantly emphasize concerns of non-antibiotic human-targeted pharmaceuticals for enhancing the spread of antibiotic resistance.

Research highlights: antibiotic resistance genes: from wastewater into the environment

2015 ◽  
Vol 1 (3) ◽  
pp. 264-267 ◽  
Author(s):  
David T. Tan ◽  
Danmeng Shuai
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Natural Transformation ◽  
Antibiotic Resistance Genes ◽  
Quantitative Model ◽  
Land Application ◽  
Untreated Wastewater

We highlight the effects of treated and untreated wastewater on antibiotic resistance genes (ARGs) in the environment, attenuation of ARGs following land application of wastewater solids, and a quantitative model for natural transformation.

scholarly journals Horizontal gene transfer facilitates the spread of extracellular antibiotic resistance genes in soil

2021 ◽  
Author(s):  
Heather A. Kittredge ◽  
Kevin M. Dougherty ◽  
Sarah E. Evans
Keyword(s):  
Antibiotic Resistance ◽  
Gene Transfer ◽  
Horizontal Gene Transfer ◽  
Resistance Genes ◽  
Natural Transformation ◽  
Antibiotic Resistance Genes ◽  
Live Cells ◽  
Resistant Bacteria ◽  
Antibiotic Resistant Bacteria ◽  
Antibiotic Resistant

AbstractAntibiotic resistance genes (ARGs) are ubiquitous in the environment and pose a serious risk to human and veterinary health. While many studies focus on the spread of live antibiotic resistant bacteria throughout the environment, it is unclear whether extracellular ARGs from dead cells can transfer to live bacteria to facilitate the evolution of antibiotic resistance in nature. Here, we inoculate antibiotic-free soil with extracellular ARGs (eARGs) from dead Pseudeononas stutzeri cells and track the evolution of antibiotic resistance via natural transformation – a mechanism of horizontal gene transfer involving the genomic integration of eARGs. We find that transformation facilitates the rapid evolution of antibiotic resistance even when eARGs occur at low concentrations (0.25 μg g-1 soil). However, when eARGs are abundant, transformation increases substantially. The evolution of antibiotic resistance was high under soil moistures typical in terrestrial systems (5%-30% gravimetric water content) and was only inhibited at very high soil moistures (>30%). While eARGs transformed into live cells at a low frequency, exposure to a low dose of antibiotic allowed a small number of transformants to reach high abundances in laboratory populations, suggesting even rare transformation events pose a risk to human health. Overall, this work demonstrates that dead bacteria and their eARGs are an overlooked path to antibiotic resistance, and that disinfection alone is insufficient to stop the spread of antibiotic resistance. More generally, the spread of eARGs in antibiotic-free soil suggests that transformation allows genetic variants to establish at low frequencies in the absence of antibiotic selection.ImportanceOver the last decade, antibiotics in the environment have gained increasing attention because they can select for drug-resistant phenotypes that would have otherwise gone extinct. To counter this effect, bacterial populations exposed to antibiotics often undergo disinfection. However, the release of extracellular antibiotic resistance genes (eARGs) into the environment following disinfection can promote the transfer of eARGs through natural transformation. This phenomenon is well-documented in wastewater and drinking water, but yet to be investigated in soil. Our results directly demonstrate that eARGs from dead bacteria are an important, but often overlooked source of antibiotic resistance in soil. We conclude that disinfection alone is insufficient to prevent the spread of ARGs. Special caution should be taken in releasing antibiotics into the environment, even if there are no live antibiotic resistant bacteria in the community, as transformation allows DNA to maintain its biological activity past microbial death.

scholarly journals Nonnutritive sweeteners can promote the dissemination of antibiotic resistance through conjugative gene transfer

2021 ◽  
Author(s):  
Zhigang Yu ◽  
Yue Wang ◽  
Ji Lu ◽  
Philip L. Bond ◽  
Jianhua Guo
Keyword(s):  
Antibiotic Resistance ◽  
Gene Transfer ◽  
Cell Membrane ◽  
Membrane Permeability ◽  
Resistance Genes ◽  
Food Additives ◽  
Antibiotic Resistance Genes ◽  
Sos Response ◽  
Conjugative Plasmid ◽  
Cell Membrane Permeability

AbstractAntimicrobial resistance (AMR) poses a worldwide threat to human health and biosecurity. The spread of antibiotic resistance genes (ARGs) via conjugative plasmid transfer is a major contributor to the evolution of this resistance. Although permitted as safe food additives, compounds such as saccharine, sucralose, aspartame, and acesulfame potassium that are commonly used as nonnutritive sweeteners have recently been associated with shifts in the gut microbiota similar to those caused by antibiotics. As antibiotics can promote the spread of antibiotic resistance genes (ARGs), we hypothesize that these nonnutritive sweeteners could have a similar effect. Here, we demonstrate for the first time that saccharine, sucralose, aspartame, and acesulfame potassium could promote plasmid-mediated conjugative transfer in three established conjugation models between the same and different phylogenetic strains. The real-time dynamic conjugation process was visualized at the single-cell level. Bacteria exposed to the tested compounds exhibited increased reactive oxygen species (ROS) production, the SOS response, and gene transfer. In addition, cell membrane permeability increased in both parental bacteria under exposure to the tested compounds. The expression of genes involved in ROS detoxification, the SOS response, and cell membrane permeability was significantly upregulated under sweetener treatment. In conclusion, exposure to nonnutritive sweeteners enhances conjugation in bacteria. Our findings provide insight into AMR spread and indicate the potential risk associated with the presence of nonnutritive sweeteners.

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