scholarly journals Improved Electrotransformation and Decreased Antibiotic Resistance of the Cystic Fibrosis Pathogen Burkholderia cenocepacia Strain J2315

2009 ◽  
Vol 76 (4) ◽  
pp. 1095-1102 ◽  
Author(s):  
Nelly Dubarry ◽  
Wenli Du ◽  
David Lane ◽  
Franck Pasta
Keyword(s):  
Cystic Fibrosis ◽  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Efflux Pump ◽  
Antibiotic Resistance Genes ◽  
Burkholderia Cenocepacia ◽  
The Poor ◽  
High Level

ABSTRACT The bacterium Burkholderia cenocepacia is pathogenic for sufferers from cystic fibrosis (CF) and certain immunocompromised conditions. The B. cenocepacia strain most frequently isolated from CF patients, and which serves as the reference for CF epidemiology, is J2315. The J2315 genome is split into three chromosomes and one plasmid. The strain was sequenced several years ago, and its annotation has been released recently. This information should allow genetic experimentation with J2315, but two major impediments appear: the poor potential of J2315 to act as a recipient in transformation and conjugation and the high level of resistance it mounts to nearly all antibiotics. Here, we describe modifications to the standard electroporation procedure that allow routine transformation of J2315 by DNA. In addition, we show that deletion of an efflux pump gene and addition of spermine to the medium enhance the sensitivity of J2315 to certain commonly used antibiotics and so allow a wider range of antibiotic resistance genes to be used for selection.

scholarly journals Novel Soil-Derived Beta-Lactam, Chloramphenicol, Fosfomycin and Trimethoprim Resistance Genes Revealed by Functional Metagenomics

Antibiotics ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 378
Author(s):  
Inka Marie Willms ◽  
Maja Grote ◽  
Melissa Kocatürk ◽  
Lukas Singhoff ◽  
Alina Andrea Kraft ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
Reference Database ◽  
Functional Metagenomics ◽  
Beta Lactam ◽  
Public Attention ◽  
Soil Ecosystems ◽  
High Level ◽  
Antibiotic Efflux

Antibiotic resistance genes (ARGs) in soil are considered to represent one of the largest environmental resistomes on our planet. As these genes can potentially be disseminated among microorganisms via horizontal gene transfer (HGT) and in some cases are acquired by clinical pathogens, knowledge about their diversity, mobility and encoded resistance spectra gained increasing public attention. This knowledge offers opportunities with respect to improved risk prediction and development of strategies to tackle antibiotic resistance, and might help to direct the design of novel antibiotics, before further resistances reach hospital settings or the animal sector. Here, metagenomic libraries, which comprise genes of cultivated microorganisms, but, importantly, also those carried by the uncultured microbial majority, were screened for novel ARGs from forest and grassland soils. We detected three new beta-lactam, a so far unknown chloramphenicol, a novel fosfomycin, as well as three previously undiscovered trimethoprim resistance genes. These ARGs were derived from phylogenetically diverse soil bacteria and predicted to encode antibiotic inactivation, antibiotic efflux, or alternative variants of target enzymes. Moreover, deduced gene products show a minimum identity of ~21% to reference database entries and confer high-level resistance. This highlights the vast potential of functional metagenomics for the discovery of novel ARGs from soil ecosystems.

scholarly journals Correlation between the Antibiotic Resistance Genes and Susceptibility to Antibiotics among the Carbapenem-Resistant Gram-Negative Pathogens

Antibiotics ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 255
Author(s):  
Salma M. Abdelaziz ◽  
Khaled M. Aboshanab ◽  
Ibrahim S. Yahia ◽  
Mahmoud A. Yassien ◽  
Nadia A. Hassouna
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Efflux Pump ◽  
Antibiotic Resistance Genes ◽  
Multiple Drug ◽  
P Value ◽  
Multidrug Efflux ◽  
Multidrug Efflux Pump ◽  
Gram Negative ◽  
Carbapenem Resistant

In this study, the correlation between the antibiotic resistance genes and antibiotic susceptibility among the carbapenem-resistant Gram-negative pathogens (CRGNPs) recovered from patients diagnosed with acute pneumonia in Egypt was found. A total of 194 isolates including Klebsiella pneumoniae (89; 46%), Escherichia coli (47; 24%) and Pseudomonas aeruginosa (58; 30%) were recovered. Of these, 34 (18%) isolates were multiple drug resistant (MDR) and carbapenem resistant. For the K. pneumoniae MDR isolates (n = 22), blaNDM (14; 64%) was the most prevalent carbapenemase, followed by blaOXA-48 (11; 50%) and blaVIM (4; 18%). A significant association (p value < 0.05) was observed between the multidrug efflux pump (AcrA) and resistance to β-lactams and the aminoglycoside acetyl transferase gene (aac-6’-Ib) gene and resistance to ciprofloxacin, azithromycin and β-lactams (except for aztreonam). For P. aeruginosa, a significant association was noticed between the presence of the blaSHV gene and the multidrug efflux pump (MexA) and resistance to fluoroquinolones, amikacin, tobramycin, co-trimoxazole and β-lactams and between the aac-6’-Ib gene and resistance to aminoglycosides. All P. aeruginosa isolates (100%) harbored the MexAB-OprM multidrug efflux pump while 86% of the K. pneumoniae isolates harbored the AcrAB-TolC pump. Our results are of great medical importance for the guidance of healthcare practitioners for effective antibiotic prescription.

Ultraviolet irradiation sensitizes Pseudomonas aeruginosa PAO1 to multiple antibiotics

2018 ◽  
Vol 4 (12) ◽  
pp. 2051-2057 ◽  
Author(s):  
Fuzheng Zhao ◽  
Qing Hu ◽  
Hongqiang Ren ◽  
Xu-Xiang Zhang
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Antibiotic Resistance ◽  
Uv Irradiation ◽  
Resistance Genes ◽  
Ultraviolet Irradiation ◽  
Efflux Pump ◽  
Antibiotic Resistance Genes ◽  
Regulatory System ◽  
Pseudomonas Aeruginosa Pao1 ◽  
Multiple Antibiotics

UV irradiation disturbs the regulatory system of efflux pump proteins to sensitize P. aeruginosa to multiple antibiotics. The increasing susceptibility to rifampicin and vancomycin might be caused by UV-mediated mutations in antibiotic resistance genes.

scholarly journals Draft Genome Sequences of Five Pseudomonas aeruginosa Clinical Strains Isolated from Sputum Samples from Cystic Fibrosis Patients: TABLE 1

2016 ◽  
Vol 4 (1) ◽  
Author(s):  
M. B. Couger ◽  
Anna Wright ◽  
Erika I. Lutter ◽  
Noha Youssef
Keyword(s):  
Cystic Fibrosis ◽  
Pseudomonas Aeruginosa ◽  
Antibiotic Resistance ◽  
Virulence Factors ◽  
Resistance Genes ◽  
Draft Genome ◽  
Antibiotic Resistance Genes ◽  
Genome Sequences ◽  
Clinical Strains ◽  
Chronic Colonization

We report here the draft genome sequences of five Pseudomonas aeruginosa isolates obtained from sputum samples from two cystic fibrosis patients with chronic colonization. These closely related strains harbor 225 to 493 genes absent from the P. aeruginosa POA1 genome and contain 178 to 179 virulence factors and 29 to 31 antibiotic resistance genes.

scholarly journals Cotransfer of Antibiotic Resistance Genes and a hylEfm-Containing Virulence Plasmid in Enterococcus faecium

2009 ◽  
Vol 53 (10) ◽  
pp. 4240-4246 ◽  
Author(s):  
Cesar A. Arias ◽  
Diana Panesso ◽  
Kavindra V. Singh ◽  
Louis B. Rice ◽  
Barbara E. Murray
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Enterococcus Faecium ◽  
Antibiotic Resistance Genes ◽  
The United States ◽  
Virulence Plasmid ◽  
Gene Encoding ◽  
Healthy Human ◽  
Gentamicin Resistance ◽  
High Level

ABSTRACT The hylEfm gene (encoding a putative hyaluronidase) has been found almost exclusively in Enterococcus faecium clinical isolates, and recently, it was shown to be on a plasmid which increased the ability of E. faecium strains to colonize the gastrointestinal tract. In this work, the results of mating experiments between hylEfm -containing strains of E. faecium belonging to clonal cluster 17 and isolated in the United States and Colombia indicated that the hylEfm gene of these strains is also carried on large plasmids (>145 kb) which we showed transfer readily from clinical strains to E. faecium hosts. Cotransfer of resistance to vancomycin and high-level resistance (HLR) to aminoglycosides (gentamicin and streptomycin) and erythromycin was also observed. The vanA gene cluster and gentamicin resistance determinants were genetically linked to hylEfm , whereas erm(B) and ant(6)-I, conferring macrolide-lincosamide-streptogramin B resistance and HLR to streptomycin, respectively, were not. A hylEfm -positive transconjugant resulting from a mating between a well-characterized endocarditis strain [TX0016 (DO)] and a derivative of a fecal strain of E. faecium from a healthy human volunteer (TX1330RF) exhibited increased virulence in a mouse peritonitis model. These results indicate that E. faecium strains use a strategy which involves the recruitment into the same genetic unit of antibiotic resistance genes and determinants that increase the ability to produce disease. Our findings indicate that the acquisition of the hylEfm plasmids may explain, at least in part, the recent successful emergence of some E. faecium strains as nosocomial pathogens.

scholarly journals Screening and Quantification of the Expression of Antibiotic Resistance Genes in Acinetobacter baumannii with a Microarray

2009 ◽  
Vol 54 (1) ◽  
pp. 333-340 ◽  
Author(s):  
Sébastien Coyne ◽  
Ghislaine Guigon ◽  
Patrice Courvalin ◽  
Bruno Périchon
Keyword(s):  
Antibiotic Resistance ◽  
Acinetobacter Baumannii ◽  
Resistance Genes ◽  
Efflux Pump ◽  
Acquired Resistance ◽  
Antibiotic Resistance Genes ◽  
Efflux Pumps ◽  
Single Step ◽  
Resistance Determinants

ABSTRACT An oligonucleotide-based DNA microarray was developed to evaluate expression of genes for efflux pumps in Acinetobacter baumannii and to detect acquired antibiotic resistance determinants. The microarray contained probes for 205 genes, including those for 47 efflux systems, 55 resistance determinants, and 35 housekeeping genes. The microarray was validated by comparative analysis of mutants overexpressing or deficient in the pumps relative to the parental strain. The performance of the microarray was also evaluated using in vitro single-step mutants obtained on various antibiotics. Overexpression, confirmed by quantitative reverse transcriptase PCR, of RND efflux pumps AdeABC, due to a G30D substitution in AdeS in a multidrug-resistant (MDR) strain obtained on gentamicin, and AdeIJK, in two mutants obtained on cefotaxime or tetracycline, was detected. A new efflux pump, AdeFGH, was found to be overexpressed in a mutant obtained on chloramphenicol. Study of MDR clinical isolates, including the AYE strain, whose entire sequence has been determined, indicated overexpression of AdeABC and of the chromosomally encoded cephalosporinase as well as the presence of several acquired resistance genes. The overexpressed and acquired determinants detected by the microarray could account for nearly the entire MDR phenotype of the isolates. The microarray is potentially useful for detection of resistance in A. baumannii and should allow detection of new efflux systems associated with antibiotic resistance.

scholarly journals Induction of Multidrug Resistance Mechanism in Escherichia coli Biofilms by Interplay between Tetracycline and Ampicillin Resistance Genes

2009 ◽  
Vol 53 (11) ◽  
pp. 4628-4639 ◽  
Author(s):  
Thithiwat May ◽  
Akinobu Ito ◽  
Satoshi Okabe
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Antimicrobial Agents ◽  
Efflux Pump ◽  
Marker Genes ◽  
Ampicillin Resistance ◽  
High Level ◽  
Biofilm Development ◽  
Level Resistance

ABSTRACT Biofilms gain resistance to various antimicrobial agents, and the presence of antibiotic resistance genes is thought to contribute to a biofilm-mediated antibiotic resistance. Here we showed the interplay between the tetracycline resistance efflux pump TetA(C) and the ampicillin resistance gene (bla TEM-1) in biofilms of Escherichia coli harboring pBR322 in the presence of the mixture of ampicillin and tetracycline. E. coli in the biofilms could obtain the high-level resistance to ampicillin, tetracycline, penicillin, erythromycin, and chloramphenicol during biofilm development and maturation as a result of the interplay between the marker genes on the plasmids, the increase of plasmid copy number, and consequently the induction of the efflux systems on the bacterial chromosome, especially the EmrY/K and EvgA/S pumps. In addition, we characterized the overexpression of the TetA(C) pump that contributed to osmotic stress response and was involved in the induction of capsular colanic acid production, promoting formation of mature biofilms. However, this investigated phenomenon was highly dependent on the addition of the subinhibitory concentrations of antibiotic mixture, and the biofilm resistance behavior was limited to aminoglycoside antibiotics. Thus, marker genes on plasmids played an important role in both resistance of biofilm cells to antibiotics and in formation of mature biofilms, as they could trigger specific chromosomal resistance mechanisms to confer a high-level resistance during biofilm formation.

scholarly journals Diversity and Distribution of Potential Pathogens and Antibiotic Resistance Genes in Anthropogenic Disturbances Aquatic Environment and Their Relationship with Microbial Indicators

2021 ◽  
Author(s):  
Xiang-Long Zhao ◽  
Zhao Qi ◽  
Hao Huang ◽  
Jian Tu ◽  
Xiang-Jun Song ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Efflux Pump ◽  
Antibiotic Resistance Genes ◽  
Chaohu Lake ◽  
Lake Basin ◽  
Metagenomic Sequencing ◽  
Sequencing Data ◽  
Potential Health ◽  
Microbial Indicators

Abstract Microbial indicators are often used as alternative indicators of microbial safety in water. However, information regarding the correlation between microbial indicators and ecotoxicological factors such as potential pathogens and antibiotic resistance genes (ARGs) in anthropogenically impacted waters remains highly limited. Combining 16S rRNA and metagenomic sequencing data, we investigated the composition of bacterial community and potential pathogens, ARGs diversity, ARGs host and horizontal gene transfer (HGT) potential in water samples under the influence of different exogenous pollutants in Chaohu Lake basin. The water body that receives a large amount of domestic sewage showed a significant decrease in microbial diversity and a significant enrichment of potential pathogens. A total of 14 main types and 461 subtypes of ARGs were detected in all samples, dominated by multidrug resistance (MDR) efflux pump (53.6%), aminoglycoside (6.0%), fluoroquinolone (5.8%) and polymyxin (5.46%). Host-tracking analysis showed that Escherichia coli and Bacteroides graminisolvens carried a wealth of ARG subtypes. Correlation analysis showed that potential pathogens and some ARG subtypes such as dfrE, sul2, PmrE exhibits significant correlation with indicator bacteria. Overall, next-generation sequencing (NGS) has the ability to conduct preliminary surveys of environmental samples to access potential health risks, thus providing ideas for water resources management.

scholarly journals Detection of Variants of the pRAS3, pAB5S9, and pSN254 Plasmids in Aeromonas salmonicida subsp. salmonicida: Multidrug Resistance, Interspecies Exchanges, and Plasmid Reshaping

2014 ◽  
Vol 58 (12) ◽  
pp. 7367-7374 ◽  
Author(s):  
Antony T. Vincent ◽  
Mélanie V. Trudel ◽  
Valérie E. Paquet ◽  
Brian Boyle ◽  
Katherine H. Tanaka ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Multidrug Resistance ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
Aeromonas Salmonicida ◽  
Fish Farms ◽  
Antibiotic Resistant ◽  
Content Type ◽  
Resistance Plasmids ◽  
High Level

ABSTRACTThe ubiquitous water-borne Gram-negative bacteriumAeromonas salmonicidasubsp.salmonicidais the causative agent of furunculosis, a worldwide disease in fish farms. Plasmids carrying antibiotic resistance genes have already been described for this bacterium. The aim of the present study was to identify and characterize additional multidrug resistance plasmids inA. salmonicidasubsp.salmonicida. We sequenced the plasmids present in two multiple antibiotic-resistant isolates using high-throughput technologies. We also investigated 19 other isolates with various multidrug resistance profiles by genotyping PCR and assessed their resistance to tetracycline. We identified variants of the pAB5S9 and pSN254 plasmids that carry several antibiotic resistance genes and that have been previously reported in bacteria other thanA. salmonicidasubsp.salmonicida, which suggests a high level of interspecies exchange. Genotyping analyses and the antibiotic resistance profiles of the 19 other isolates support the idea that multiple versions of pAB5S9 and pSN254 exist inA. salmonicidasubsp.salmonicida. We also identified variants of the pRAS3 plasmid. The present study revealed thatA. salmonicidasubsp.salmonicidaharbors a wide variety of plasmids, which suggests that this ubiquitous bacterium may contribute to the spread of antibiotic resistance genes in the environment.

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