scholarly journals Conjugation factors controlling F-plasmid antibiotic resistance transmission

2018 ◽  
Author(s):  
Hanna Alalam ◽  
Fabrice E. Graf ◽  
Martin Palm ◽  
Marie Abadikhah ◽  
Martin Zackrisson ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Horizontal Transmission ◽  
Antibiotic Resistance Genes ◽  
Host Cells ◽  
The Novel ◽  
F Plasmid ◽  
Resistance Development ◽  
E Coli ◽  
Cost Efficient ◽  
Efficient Screening

ABSTRACTThe rapid horizontal transmission of many antibiotic resistance genes between bacterial host cells on conjugative plasmids is a major cause of the accelerating antibiotic resistance crisis. Preventing understanding and targeting conjugation, there currently are no experimental platforms for fast and cost-efficient screening of genetic effects on antibiotic resistance transmission by conjugation. We introduce a novel experimental framework to screen for conjugation based horizontal transmission of antibiotic resistance between >60.000 pairs of cell populations in parallel. Plasmid-carrying donor strains are constructed in high throughput. We then mix the resistance plasmid carrying donors with recipients in a design where only transconjugants can reproduce, measure growth in dense intervals and extract transmission times as the growth lag. As proof-of-principle, we exhaustively explored chromosomal genes controlling F plasmid donation withinE. colipopulations, by screening the Keio deletion collection at high replication. We recover all six known chromosomal gene mutants affecting conjugation and identify >50 novel factors, all of which diminish antibiotic resistance transmission. We verify 10 of the novel genes’ effects in a liquid mating assay. The new framework holds great potential for exhaustive disclosing of candidate targets for helper drugs that delay resistance development in patients and societies and improves the longevity of current and future antibiotics.

scholarly journals A High-Throughput Method for Screening for Genes Controlling Bacterial Conjugation of Antibiotic Resistance

mSystems ◽  
2020 ◽  
Vol 5 (6) ◽  
pp. e01226-20
Author(s):  
Hanna Alalam ◽  
Fabrice E. Graf ◽  
Martin Palm ◽  
Marie Abadikhah ◽  
Martin Zackrisson ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Horizontal Transmission ◽  
Antibiotic Resistance Genes ◽  
Host Cells ◽  
The Novel ◽  
Content Type ◽  
Rapid Construction ◽  
Cost Efficient ◽  
Efficient Screening ◽  
New Framework

ABSTRACTThe rapid horizontal transmission of antibiotic resistance genes on conjugative plasmids between bacterial host cells is a major cause of the accelerating antibiotic resistance crisis. There are currently no experimental platforms for fast and cost-efficient screening of genetic effects on antibiotic resistance transmission by conjugation, which prevents understanding and targeting conjugation. We introduce a novel experimental framework to screen for conjugation-based horizontal transmission of antibiotic resistance between >60,000 pairs of cell populations in parallel. Plasmid-carrying donor strains are constructed in high-throughput. We then mix the resistance plasmid-carrying donors with recipients in a design where only transconjugants can reproduce, measure growth in dense intervals, and extract transmission times as the growth lag. As proof-of-principle, we exhaustively explore chromosomal genes controlling F-plasmid donation within Escherichia coli populations, by screening the Keio deletion collection in high replication. We recover all seven known chromosomal gene mutants affecting conjugation as donors and identify many novel mutants, all of which diminish antibiotic resistance transmission. We validate nine of the novel genes’ effects in liquid mating assays and complement one of the novel genes’ effect on conjugation (rseA). The new framework holds great potential for exhaustive disclosing of candidate targets for helper drugs that delay resistance development in patients and societies and improve the longevity of current and future antibiotics. Further, the platform can easily be adapted to explore interspecies conjugation, plasmid-borne factors, and experimental evolution and be used for rapid construction of strains.IMPORTANCE The rapid transmission of antibiotic resistance genes on conjugative plasmids between bacterial host cells is a major cause of the accelerating antibiotic resistance crisis. There are currently no experimental platforms for fast and cost-efficient screening of genetic effects on antibiotic resistance transmission by conjugation, which prevents understanding and targeting conjugation. We introduce a novel experimental framework to screen for conjugation-based horizontal transmission of antibiotic resistance between >60,000 pairs of cell populations in parallel. As proof-of-principle, we exhaustively explore chromosomal genes controlling F-plasmid donation within E. coli populations. We recover all previously known and many novel chromosomal gene mutants that affect conjugation efficiency. The new framework holds great potential for rapid screening of compounds that decrease transmission. Further, the platform can easily be adapted to explore interspecies conjugation, plasmid-borne factors, and experimental evolution and be used for rapid construction of strains.

scholarly journals Freshwater Viral Metagenome Reveals Novel and Functional Phage-borne Antibiotic Resistance Genes

2019 ◽  
Author(s):  
Kira Moon ◽  
Ilnam Kang ◽  
Kwang Seung Park ◽  
Jeong Ho Jeon ◽  
Kihyun lee ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
Viral Metagenomics ◽  
The Novel ◽  
Multidrug Efflux ◽  
Bacterial Populations ◽  
Host Interactions ◽  
E Coli ◽  
Catalytic Kinetics

Abstract BackgroundAntibiotic resistance developed by bacteria is a significant threat to global health. Antibiotic resistance genes (ARGs) spread across different bacterial populations through multiple dissemination routes, including horizontal gene transfer mediated by bacteriophages. ARGs carried by bacteriophages are considered especially threatening due to their prolonged persistence in the environment, fast replication rates, and ability to infect phylogenetically remote bacterial hosts. Several studies employing qPCR and viral metagenomics have shown that viral fraction and viral sequence reads in clinical and environmental samples carry many ARGs. However, only a few ARGs have been found in viral contigs assembled from metagenome reads, with most of these genes lacking effective antibiotic resistance phenotypes. Owing to the wide application of viral metagenomics, nevertheless, different classes of ARGs are being continuously found in viral metagenomes acquired from diverse environments. As such, the presence and functionality of ARGs encoded by bacteriophages remain up for debate.ResultsWe evaluated ARGs excavated from viral contigs recovered from urban surface water viral metagenome data. In virome reads and contigs, diverse ARGs, including polymyxin resistance genes, multidrug efflux proteins, and β-lactamases, were identified. In particular, the novel β-lactamases blaHRV-1 and blaHRVM-1 found in this study had unique sequences, forming distinct clades of Class A and subclass B3 β-lactamases, respectively. Minimum inhibitory concentration analyses for E. coli strains harboring blaHRV-1 and blaHRVM-1 and catalytic kinetics of purified HRV-1 and HRVM-1 showed reduced susceptibility to penicillin, narrow- and extended-spectrum cephalosporins, and carbapenems. These genes were also found in bacterial metagenomes, indicating that they were harbored by actively infecting phages.ConclusionOur results showed that viruses in the environment carry as-yet-unreported functional ARGs, albeit in small quantities. We thereby suggest that environmental bacteriophages could be reservoirs of widely variable, unknown ARGs that could be disseminated via virus-host interactions.

scholarly journals Characterization of Tn7-like transposons and its related antibiotic resistance among Enterobacteriaceae from livestock and poultry in China

2020 ◽  
Author(s):  
Juan He ◽  
Cui Li ◽  
Pengfei Cui ◽  
Hongning Wang
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Horizontal Transmission ◽  
Antibiotic Resistance Genes ◽  
Variable Region ◽  
Multidrug Resistant ◽  
Antibiotic Resistant ◽  
E Coli ◽  
New Type

Abstract Background: This study was aimed to investigate the prevalence and structure of Tn7-like in Enterobacteriaceae from livestock and poultry as well as their possible role as reservoir of antibiotic resistance genes (ARGs).Methods: Polymerase chain reaction (PCR) and DNA sequencing analyses were used for the characterization of Tn7-like, associated integrons and ARGs. The antimicrobial resistance profile of the isolates was examined by using disc diffusion test.Results: Three hundred and seventy-eight Tn7-like-positive strains of Enterobacteriaceae were isolated, and included E. coli (128), Proteus(150), K. pneumonia(17), Salmonella(13), M. morganii (21) and A. baumannii(1), wherein high resistance was observed for Trimethoprim/Sulfamethoxazole and Streptomycin, and fifty percent of the strains were multidrug-resistant. Integrons class 2 were detected in all of the isolates and there are high frequency mutation sites especially in 535, a stop mutation. Variable region of class 2 integrons carried same gene cassettes, namely aadA1-sat2-dfrA1. From the 378 isolated strains, we found a new type of Tn7-like on a plasmid, named Tn6765.Conclusions: These findings proved that the Tn7-like can contribute to the horizontal transmission of antibiotic resistant genes in livestock and poultry. As potential vessels for antibiotic resistance genes (ARGs), Tn7-like could not be ignored due to their efficient transfer ability in environments.

scholarly journals Plasmid Identification and Plasmid-Mediated Antimicrobial Gene Detection in Norwegian Isolates

2020 ◽  
Vol 9 (1) ◽  
pp. 52
Author(s):  
Abdolrahman Khezri ◽  
Ekaterina Avershina ◽  
Rafi Ahmad
Keyword(s):  
Antibiotic Resistance ◽  
Horizontal Transmission ◽  
Antibiotic Resistance Genes ◽  
Bioinformatic Analysis ◽  
Whole Genome Sequencing Data ◽  
Whole Genome ◽  
Hybrid Assembly ◽  
Sequencing Data ◽  
Gene Detection ◽  
E Coli

Norway is known for being one of the countries with the lowest levels of antimicrobial resistance (AMR). AMR, through acquired genes located on transposons or conjugative plasmids, is the horizontal transmission of genes required for a given bacteria to withstand antibiotics. In this work, bioinformatic analysis of whole-genome sequences and hybrid assembled data from Escherichia coli, and Klebsiella pneumoniae isolates from Norwegian patients was performed. For detection of putative plasmids in isolates, the plasmid assembly mode in SPAdes was used, followed by annotation of resulting contigs using PlasmidFinder and two curated plasmid databases (Brooks and PLSDB). Furthermore, ResFinder and Comprehensive Antibiotic Resistance Database (CARD) were used for the identification of antibiotic resistance genes (ARGs). The IncFIB plasmid was detected as the most prevalent plasmid in both E. coli, and K. pneumoniae isolates. Furthermore, ARGs such as aph(3″)-Ib, aph(6)-Id, sul1, sul2, tet(D), and qnrS1 were identified as the most abundant plasmid-mediated ARGs in Norwegian E. coli and K. pneumoniae isolates, respectively. Using hybrid assembly, we were able to locate plasmids and predict ARGs more confidently. In conclusion, plasmid identification and ARG detection using whole-genome sequencing data are heavily dependent on the database of choice; therefore, it is best to use several tools and/or hybrid assembly for obtaining reliable identification results.

scholarly journals Nitric oxide (NO) elicits aminoglycoside tolerance in Escherichia coli but antibiotic resistance gene carriage and NO sensitivity have not co-evolved

2021 ◽  
Author(s):  
Cláudia A. Ribeiro ◽  
Luke A. Rahman ◽  
Louis G. Holmes ◽  
Ayrianna M. Woody ◽  
Calum M. Webster ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Antibiotic Resistance ◽  
Antibiotic Susceptibility ◽  
Bacterial Pathogens ◽  
Antibiotic Resistance Gene ◽  
Antibiotic Resistance Genes ◽  
Resistance Mechanisms ◽  
Current Work ◽  
E Coli ◽  
Respiratory Inhibitor

AbstractThe spread of multidrug-resistance in Gram-negative bacterial pathogens presents a major clinical challenge, and new approaches are required to combat these organisms. Nitric oxide (NO) is a well-known antimicrobial that is produced by the immune system in response to infection, and numerous studies have demonstrated that NO is a respiratory inhibitor with both bacteriostatic and bactericidal properties. However, given that loss of aerobic respiratory complexes is known to diminish antibiotic efficacy, it was hypothesised that the potent respiratory inhibitor NO would elicit similar effects. Indeed, the current work demonstrates that pre-exposure to NO-releasers elicits a > tenfold increase in IC50 for gentamicin against pathogenic E. coli (i.e. a huge decrease in lethality). It was therefore hypothesised that hyper-sensitivity to NO may have arisen in bacterial pathogens and that this trait could promote the acquisition of antibiotic-resistance mechanisms through enabling cells to persist in the presence of toxic levels of antibiotic. To test this hypothesis, genomics and microbiological approaches were used to screen a collection of E. coli clinical isolates for antibiotic susceptibility and NO tolerance, although the data did not support a correlation between increased carriage of antibiotic resistance genes and NO tolerance. However, the current work has important implications for how antibiotic susceptibility might be measured in future (i.e. ± NO) and underlines the evolutionary advantage for bacterial pathogens to maintain tolerance to toxic levels of NO.

scholarly journals Vibrio cholerae Triggers SOS and Mutagenesis in Response to a Wide Range of Antibiotics: a Route towards Multiresistance

2011 ◽  
Vol 55 (5) ◽  
pp. 2438-2441 ◽  
Author(s):  
Zeynep Baharoglu ◽  
Didier Mazel
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Vibrio Cholerae ◽  
Fluorescent Protein ◽  
Resistance Development ◽  
Content Type ◽  
E Coli ◽  
Wide Range ◽  
Green Fluorescent ◽  
Regulated Promoters

ABSTRACTAntibiotic resistance development has been linked to the bacterial SOS stress response. InEscherichia coli, fluoroquinolones are known to induce SOS, whereas other antibiotics, such as aminoglycosides, tetracycline, and chloramphenicol, do not. Here we address whether various antibiotics induce SOS inVibrio cholerae. Reporter green fluorescent protein (GFP) fusions were used to measure the response of SOS-regulated promoters to subinhibitory concentrations of antibiotics. We show that unlike the situation withE. coli, all these antibiotics induce SOS inV. cholerae.

scholarly journals Mechanisms of Resistance in Multiple-Antibiotic-Resistant Escherichia coli Strains of Human, Animal, and Food Origins

2004 ◽  
Vol 48 (10) ◽  
pp. 3996-4001 ◽  
Author(s):  
Yolanda Sáenz ◽  
Laura Briñas ◽  
Elena Domínguez ◽  
Joaquim Ruiz ◽  
Myriam Zarazaga ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Amino Acid ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
Mechanisms Of Resistance ◽  
Antibiotic Resistant ◽  
E Coli ◽  
Class 1 ◽  
Human Animal

ABSTRACT Seventeen multiple-antibiotic-resistant nonpathogenic Escherichia coli strains of human, animal, and food origins showed a wide variety of antibiotic resistance genes, many of them carried by class 1 and class 2 integrons. Amino acid changes in MarR and mutations in marO were identified for 15 and 14 E. coli strains, respectively.

scholarly journals Antibiotic-resistant Escherichia coli and Salmonella spp. associated with dairy cattle and farm environment having public health significance

2019 ◽  
Vol 12 (7) ◽  
pp. 984-993 ◽  
Author(s):  
Md. Abdus Sobur ◽  
Abdullah Al Momen Sabuj ◽  
Ripon Sarker ◽  
A. M. M. Taufiqur Rahman ◽  
S. M. Lutful Kabir ◽  
...  
Keyword(s):  
Public Health ◽  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Resistance Genes ◽  
One Health ◽  
Dairy Farm ◽  
Antibiotic Resistance Genes ◽  
Salmonella Spp ◽  
Antibiotic Resistant ◽  
E Coli

Aim: The present study was carried out to determine load of total bacteria, Escherichia coli and Salmonella spp. in dairy farm and its environmental components. In addition, the antibiogram profile of the isolated bacteria having public health impact was also determined along with identification of virulence and resistance genes by polymerase chain reaction (PCR) under a one-health approach. Materials and Methods: A total of 240 samples of six types (cow dung - 15, milk - 10, milkers' hand wash - 10, soil - 10 water - 5, and vegetables - 10) were collected from four dairy farms. For enumeration, the samples were cultured onto plate count agar, eosin methylene blue, and xylose-lysine deoxycholate agar and the isolation and identification of the E. coli and Salmonella spp. were performed based on morphology, cultural, staining, and biochemical properties followed by PCR. The pathogenic strains of E. coli stx1, stx2, and rfbO157 were also identified through PCR. The isolates were subjected to antimicrobial susceptibility test against 12 commonly used antibiotics by disk diffusion method. Detection of antibiotic resistance genes ereA, tetA, tetB, and SHV were performed by PCR. Results: The mean total bacterial count, E. coli and Salmonella spp. count in the samples ranged from 4.54±0.05 to 8.65±0.06, 3.62±0.07 to 7.04±0.48, and 2.52±0.08 to 5.87±0.05 log colony-forming unit/g or ml, respectively. Out of 240 samples, 180 (75%) isolates of E. coli and 136 (56.67%) isolates of Salmonella spp. were recovered through cultural and molecular tests. Among the 180 E. coli isolates, 47 (26.11%) were found positive for the presence of all the three virulent genes, of which stx1 was the most prevalent (13.33%). Only three isolates were identified as enterohemorrhagic E. coli. Antibiotic sensitivity test revealed that both E. coli and Salmonella spp. were found highly resistant to azithromycin, tetracycline, erythromycin, oxytetracycline, and ertapenem and susceptible to gentamycin, ciprofloxacin, and imipenem. Among the four antibiotic resistance genes, the most observable was tetA (80.51-84.74%) in E. coli and Salmonella spp. and SHV genes were the lowest one (22.06-25%). Conclusion: Dairy farm and their environmental components carry antibiotic-resistant pathogenic E. coli and Salmonella spp. that are potential threat for human health which requires a one-health approach to combat the threat.

scholarly journals Microbiological monitoring as a component of efficient prevention and treatment of purulent-septic infections in an orthopedics and traumatology department

2021 ◽  
Vol 23 (3) ◽  
pp. 381-387
Author(s):  
N. M. Polishchuk ◽  
D. L. Kyryk ◽  
I. Ye. Yurchuk
Keyword(s):  
Antibiotic Resistance ◽  
Emergency Care ◽  
Antibiotic Susceptibility ◽  
Analytical Data ◽  
Empirical Therapy ◽  
Care Hospital ◽  
Resistance Development ◽  
E Coli ◽  
Microbiological Monitoring ◽  
Bacteriological Test

Efficient monitoring of circulating purulent-septic infectious agents in a clinical setting and a study on antibiotic susceptibility of isolated strains of microorganisms allows identifying changes in the pathogen structure and trends in antibiotic resistance development, which helps to determine the tactics of antibacterial therapy and elaborate appropriate measures. The aim of the study. Retrospective analysis of the results of microbiological monitoring of purulent-septic infectious (PSI) agents in the Orthopedics and Traumatology Department (OTD) of the Zaporizhzhia Central Ambulance and Emergency Care Hospital over the period 2017–2020 to determine the main antibacterial drugs for empirical therapy. Materials and methods. We analyzed the bacteriological test results of 664 clinical material samples obtained from OTD patients using bacteriological examination statistical reporting and analytical data of the WHONET 5.6 software. Results. The main PSI pathogens in the OTD were from the ESKAPE group: E. coli, S. aureus, K. pneumoniae, A. baumannii, E. faecalis, P. aeruginosa and S. epidermidis, P. mirabilis, C. amycolatum. Isolates of E. faecalis were sensitive to vancomycin, linezolid, S. aureus – to linezolid, tigecycline, netilmicin, A. baumannii – to tigecycline. All P. aeruginosa strains were resistant to ticarcillin/clavulanate, cefepime, chloramphenicol, imipenem, meropenem, aztreonam, ciprofloxacin. E. coli and K. pneumoniae were resistant to ampicillin, ticarcillin/clavulanate, aztreonam, ceftriaxone, cefepime. The number of isolates sensitive to piperacillin/tazobactam, carbapenems, levofloxacin, gentamicin, amikacin, chloramphenicol ranged from 37 % to 65 %. Conclusions. E. coli, S. aureus, K. pneumoniae, A. baumannii, E. faecalis, P. aeruginosa, S. epidermidis, P. mirabilis, C. amycolatum play an important role in the structure of PSI pathogens in the Orthopedics and Traumatology Department of Zaporizhzhia Central Ambulance and Emergency Care Hospital. The antibiotics of choice as the antibacterial empirical therapy for enterococcal infections are vancomycin, linezolid, for staphylococcal infections – vancomycin, linezolid, tigecycline, netilmicin. PSI pathogens continually evolve developing antibiotic resistance, and it is of particular importance to monitor antibiotic susceptibility of microorganisms within the OTD.

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