Year-end Sale % OFF 
Abstract
Background: Phenotypic characterization of antimicrobial resistance (AMR) in bacteria has remained the gold standard for investigation and monitoring of what resistance is present in an organism. However, the process is laborious and not attractive for screening multiple plasmids from a microbial community (plasmidomes). Instead, genomic tools are used, but a major bottle neck that presence of genes does not always translate into phenotypes. Methods: We designed the plasmidome AMR screening (PAMRS) workflow to investigate the presence of antibiotic resistant phenotypes in a plasmidome using Escherichia coli as a host organism. Plasmidomes were extracted from the faecal matter of chicken, cattle and humans using commercial plasmid extraction kits. Competent E. coli cells were transformed and evaluated using disk diffusion. Thirteen antibiotic resistant phenotypes were screened. Results: Here, we show that multiple antibiotic resistant phenotypes encoded by plasmids can be rapidly screened simultaneously using the PAMRS workflow. E. coli was able to pick up to 7, 5 or 8 resistant phenotypes from a single plasmidome from chicken, cattle or humans, respectively. Resistance to ceftazidime was the most frequently picked up phenotype in humans (52.6%) and cattle (90.5%), whereas in chickens, the most picked up resistant phenotype was resistance to co-trimoxazole, ceftriaxone and ampicillin (18.4% each). Conclusions: This workflow is a novel tool that could facilitate studies to evaluate the occurrence and expression of plasmid-encoded antibiotic resistance in microbial communities and their associated plasmid-host ranges. It could find application in the screening of plasmid-encoded virulence genes.
Highlights
Antibiotic-resistant bacterial infections are a major public health problem
Data suggest that E. coli can be transformed with extracellular plasmid DNA under natural conditions[17,18], and these observations suggest that bacteria could be transformed by DNA in the gut and may contribute to the spread of antibiotic resistance genes (ARGs)
To determine the burden of active or expressed resistance genes in human, chicken, and cattle gut microbiomes, we developed the Plasmidome antimicrobial resistance (AMR) screening (PAMRS) workflow using a combination of plasmidome isolation, bacterial transformation, and multi-disk diffusion to rapidly investigate the ecology of thirteen (13) antibiotic resistant phenotypes from gut plasmidomes using Escherichia coli as a host
Summary
Antibiotic-resistant bacterial infections are a major public health problem. Annually, some estimated 30,000 deaths are recorded in the EU alone[1], while the morbidity and mortality in low to middle income countries in South America, Asia and Africa are even higher[2,3,4]. Over 80 bacterial species can be transformed due to the presence of genes involved in DNA uptake, suggesting that, this trait of competency is widespread[14]. Methods: We designed the plasmidome AMR screening (PAMRS) workflow to investigate the presence of antibiotic resistant phenotypes in a plasmidome using Escherichia coli as a host organism. Conclusions: This workflow is a novel tool that could facilitate studies to evaluate the occurrence and expression of plasmid-encoded antibiotic resistance in microbial communities and their associated plasmid-host ranges. It could find application in the screening of Invited Reviewers 1 version 1
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
