scholarly journals Emergence of Azithromycin Resistance Mediated by Phosphotransferase-Encoding mph(A) in Diarrheagenic Vibrio fluvialis

mSphere ◽  
2019 ◽  
Vol 4 (3) ◽  
Author(s):  
Goutam Chowdhury ◽  
Thandavarayan Ramamurthy ◽  
Amit Ghosh ◽  
Shanta Dutta ◽  
Eizo Takahashi ◽  
...  
Keyword(s):  
Public Health ◽  
Antibiotic Resistance ◽  
Bacterial Species ◽  
Antibiotic Resistance Gene ◽  
Public Health Importance ◽  
Class 1 Integron ◽  
Content Type ◽  
Genetic Lineages ◽  
Vibrio Fluvialis ◽  
Azithromycin Resistance

ABSTRACT The azithromycin resistance conferred by phosphotransferase is encoded in the gene mph(A). This gene has been discovered in and reported for many bacterial species. We examined the prevalence of azithromycin resistance in Vibrio fluvialis (AR-VF) isolated during 2014 to 2015 from the hospitalized acute diarrheal patients in Kolkata, India. Most of the V. fluvialis isolates are identified as the sole pathogen (54%). The prevalence of AR-VF was higher in 2015 (19 [68%]) than in 2014 (9 [32%]). Among AR-VF isolates, the azithromycin MICs ranged from 4 to >256 mg/liter. Twenty-eight of the 48 (58%) V. fluvialis isolates harbored the gene mph(A) and phenotypically resistant to azithromycin. All the AR-VF isolates remained susceptible to doxycycline. In addition to azithromycin, other antimicrobial resistance-encoding genes of AR-VF were also characterized. All the AR-VF isolates were positive for class 1 integron, and most of them (17/28) carried the dfrA1 gene cassettes. Only one isolate was positive for the ereA gene, which encodes resistance to erythomycin. The majority of the isolates were resistant to β-lactam antibiotics (blaOXA-1 [96%], blaOXA-7 [93%], and blaTEM-9 [68%]) and aminoglycoside actetyltransferase, conferring resistance to ciprofloxacin-modifying enzyme [aac(6′)Ib-cr] (96%). Analyses by pulsed-field gel electrophoresis (PFGE) showed that the AR-VF isolates belonged to different genetic lineages. This is the first study to report azithromycin resistance and the presence of the mph(A) gene in V. fluvialis isolates. Circulation of AR-VF isolates with high azithromycin MICs is worrisome, since it may limit the treatment options for diarrheal infections. IMPORTANCE The progressive rise in antibiotic resistance among enteric pathogens in developing countries is becoming a big concern. India is one of the largest consumers of antibiotics, and their use is not well regulated. V. fluvialis is increasingly recognized as an emerging diarrheal pathogen of public health importance. Here we report the emergence of azithromycin resistance in V. fluvialis isolates from diarrheal patients in Kolkata, India. Azithromycin has been widely used in the treatment of various infections, both in children and in adults. Resistance to azithromycin is encoded in the gene mph(A). Emerging azithromycin resistance in V. fluvialis is a major public health challenge, and future studies should be focused on identifying ways to prevent the dissemination of this antibiotic resistance gene.

scholarly journals Expanding U.S. Laboratory Capacity for Neisseria gonorrhoeae Antimicrobial Susceptibility Testing and Whole-Genome Sequencing through the CDC's Antibiotic Resistance Laboratory Network

2020 ◽  
Vol 58 (4) ◽  
Author(s):  
Ellen N. Kersh ◽  
Cau D. Pham ◽  
John R. Papp ◽  
Robert Myers ◽  
Richard Steece ◽  
...  
Keyword(s):  
Public Health ◽  
Antibiotic Resistance ◽  
Neisseria Gonorrhoeae ◽  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Antimicrobial Susceptibility ◽  
Susceptibility Testing ◽  
Whole Genome ◽  
Content Type ◽  
Laboratory Capacity

ABSTRACT U.S. gonorrhea rates are rising, and antibiotic-resistant Neisseria gonorrhoeae (AR-Ng) is an urgent public health threat. Since implementation of nucleic acid amplification tests for N. gonorrhoeae identification, the capacity for culturing N. gonorrhoeae in the United States has declined, along with the ability to perform culture-based antimicrobial susceptibility testing (AST). Yet AST is critical for detecting and monitoring AR-Ng. In 2016, the CDC established the Antibiotic Resistance Laboratory Network (AR Lab Network) to shore up the national capacity for detecting several resistance threats including N. gonorrhoeae. AR-Ng testing, a subactivity of the CDC’s AR Lab Network, is performed in a tiered network of approximately 35 local laboratories, four regional laboratories (state public health laboratories in Maryland, Tennessee, Texas, and Washington), and the CDC’s national reference laboratory. Local laboratories receive specimens from approximately 60 clinics associated with the Gonococcal Isolate Surveillance Project (GISP), enhanced GISP (eGISP), and the program Strengthening the U.S. Response to Resistant Gonorrhea (SURRG). They isolate and ship up to 20,000 isolates to regional laboratories for culture-based agar dilution AST with seven antibiotics and for whole-genome sequencing of up to 5,000 isolates. The CDC further examines concerning isolates and monitors genetic AR markers. During 2017 and 2018, the network tested 8,214 and 8,628 N. gonorrhoeae isolates, respectively, and the CDC received 531 and 646 concerning isolates and 605 and 3,159 sequences, respectively. In summary, the AR Lab Network supported the laboratory capacity for N. gonorrhoeae AST and associated genetic marker detection, expanding preexisting notification and analysis systems for resistance detection. Continued, robust AST and genomic capacity can help inform national public health monitoring and intervention.

scholarly journals Synonymous lysine codon usage modification in a mobile antibiotic resistance gene similarly alters protein production in bacterial species with divergent lysine codon usage biases because it removes a duplicate AAA lysine codon

2018 ◽  
Author(s):  
Mohammed Alorabi ◽  
Aisha M. AlAmri ◽  
Yuiko Takebayashi ◽  
Kate J. Heesom ◽  
Matthew B. Avison
Keyword(s):  
Antibiotic Resistance ◽  
Codon Usage ◽  
Resistance Gene ◽  
Codon Bias ◽  
Protein Production ◽  
Bacterial Species ◽  
Antibiotic Resistance Gene ◽  
Site Directed Mutagenesis ◽  
Acinetobacter Baumanii ◽  
Highly Expressed Genes

AbstractThe mobile antibiotic resistance gene blaIMP-1 is clinically important and has a synonymous AAA:AAG lysine codon usage bias of 73:27. This bias is like that seen in experimentally determined highly expressed genes in Escherichia coli and Acinetobacter baumanii, but quite different from that seen in Pseudomonas aeruginosa (26:74 AAA:AAG). Here we show that, paradoxically, shifting the AAA:AAG lysine codon bias to 8:92 in blaIMP-1 expressed from a natural promoter results in significantly more IMP-1 production in all three species. Sequential site directed mutagenesis revealed that increased IMP-1 production occurs following removal of an AAA,AAA double lysine codon and that otherwise, lysine codon usage had no observable impact on IMP-1 production. We conclude that ribosomal slippage at this poly-adenosine region reduces efficient translation of IMP-1 and that punctuating the region with guanine reduces ribosomal slippage and increases IMP-1 production.

scholarly journals Linearmycins Activate a Two-Component Signaling System Involved in Bacterial Competition and Biofilm Morphology

2017 ◽  
Vol 199 (18) ◽  
Author(s):  
Reed M. Stubbendieck ◽  
Paul D. Straight
Keyword(s):  
Antibiotic Resistance ◽  
Abc Transporter ◽  
Bacterial Communities ◽  
Biofilm Formation ◽  
Bacterial Species ◽  
Bioactive Metabolites ◽  
Signaling System ◽  
Content Type ◽  
Two Component ◽  
Analytical Approaches

ABSTRACT Bacteria use two-component signaling systems to adapt and respond to their competitors and changing environments. For instance, competitor bacteria may produce antibiotics and other bioactive metabolites and sequester nutrients. To survive, some species of bacteria escape competition through antibiotic production, biofilm formation, or motility. Specialized metabolite production and biofilm formation are relatively well understood for bacterial species in isolation. How bacteria control these functions when competitors are present is not well studied. To address fundamental questions relating to the competitive mechanisms of different species, we have developed a model system using two species of soil bacteria, Bacillus subtilis and Streptomyces sp. strain Mg1. Using this model, we previously found that linearmycins produced by Streptomyces sp. strain Mg1 cause lysis of B. subtilis cells and degradation of colony matrix. We identified strains of B. subtilis with mutations in the two-component signaling system yfiJK operon that confer dual phenotypes of specific linearmycin resistance and biofilm morphology. We determined that expression of the ATP-binding cassette (ABC) transporter yfiLMN operon, particularly yfiM and yfiN, is necessary for biofilm morphology. Using transposon mutagenesis, we identified genes that are required for YfiLMN-mediated biofilm morphology, including several chaperones. Using transcriptional fusions, we found that YfiJ signaling is activated by linearmycins and other polyene metabolites. Finally, using a truncated YfiJ, we show that YfiJ requires its transmembrane domain to activate downstream signaling. Taken together, these results suggest coordinated dual antibiotic resistance and biofilm morphology by a single multifunctional ABC transporter promotes competitive fitness of B. subtilis. IMPORTANCE DNA sequencing approaches have revealed hitherto unexplored diversity of bacterial species in a wide variety of environments that includes the gastrointestinal tract of animals and the rhizosphere of plants. Interactions between different species in bacterial communities have impacts on our health and industry. However, many approaches currently used to study whole bacterial communities do not resolve mechanistic details of interspecies interactions, including how bacteria sense and respond to their competitors. Using a competition model, we have uncovered dual functions for a previously uncharacterized two-component signaling system involved in specific antibiotic resistance and biofilm morphology. Insights gleaned from signaling within interspecies interaction models build a more complete understanding of gene functions important for bacterial communities and will enhance community-level analytical approaches.

scholarly journals A Multispecies Cluster of VIM-1 Carbapenemase-Producing Enterobacterales Linked by a Novel, Highly Conjugative, and Broad-Host-Range IncA Plasmid Forebodes the Reemergence of VIM-1

2020 ◽  
Vol 64 (4) ◽  
Author(s):  
Gabriele Arcari ◽  
Federica Maria Di Lella ◽  
Giulia Bibbolino ◽  
Fabio Mengoni ◽  
Marzia Beccaccioli ◽  
...  
Keyword(s):  
Host Range ◽  
Bacterial Species ◽  
Klebsiella Oxytoca ◽  
Gene Cassette ◽  
University Hospital ◽  
Broad Host Range ◽  
Class 1 Integron ◽  
Content Type ◽  
Carbapenem Resistant ◽  
Class 1

ABSTRACT In this study, we investigated VIM-1-producing Escherichia coli, Klebsiella oxytoca, Klebsiella pneumoniae, Citrobacter freundii, and Enterobacter cloacae strains, isolated in 2019 during a period of active surveillance of carbapenem-resistant Enterobacterales in a large university hospital in Italy. VIM-1-producing strains colonized the gut of patients, with up to three different VIM-1-positive bacterial species isolated from a single rectal swab, but also caused bloodstream infection in one colonized patient. In the multispecies cluster, blaVIM-1 was identified in a 5-gene cassette class 1 integron, associated with several genetic determinants, including the blaSHV-12, qnrS1, and mph(A) genes, located on a highly conjugative and broad-host-range IncA plasmid. The characteristics and origin of this IncA plasmid were studied.

scholarly journals The Inactivation of Enzymes Belonging to the Central Carbon Metabolism Is a Novel Mechanism of Developing Antibiotic Resistance

mSystems ◽  
2020 ◽  
Vol 5 (3) ◽  
Author(s):  
Teresa Gil-Gil ◽  
Fernando Corona ◽  
José Luis Martínez ◽  
Alejandra Bernardini
Keyword(s):  
Antibiotic Resistance ◽  
Stenotrophomonas Maltophilia ◽  
Bacterial Species ◽  
Bacterial Metabolism ◽  
Resistant Bacteria ◽  
Uridine Diphosphate ◽  
Biosynthesis Pathway ◽  
Content Type ◽  
Peptidoglycan Biosynthesis ◽  
Central Carbon

ABSTRACT Fosfomycin is a bactericidal antibiotic, analogous to phosphoenolpyruvate, that exerts its activity by inhibiting the activity of MurA. This enzyme catalyzes the first step of peptidoglycan biosynthesis, the transfer of enolpyruvate from phosphoenolpyruvate to uridine-diphosphate-N-acetylglucosamine. Fosfomycin is increasingly being used, mainly for treating infections caused by Gram-negative multidrug-resistant bacteria. The mechanisms of mutational resistance to fosfomycin in Stenotrophomonas maltophilia, an opportunistic pathogen characterized by its low susceptibility to commonly used antibiotics, were studied in the current work. None of the mechanisms reported so far for other organisms, which include the production of fosfomycin-inactivating enzymes, target modification, induction of an alternative peptidoglycan biosynthesis pathway, and the impaired entry of the antibiotic, are involved in the acquisition of such resistance by this bacterial species. Instead, the unique cause of resistance in the mutants studied is the mutational inactivation of different enzymes belonging to the Embden-Meyerhof-Parnas central metabolism pathway. The amount of intracellular fosfomycin accumulation did not change in any of these mutants, showing that neither inactivation nor transport of the antibiotic is involved. Transcriptomic analysis also showed that the mutants did not present changes in the expression level of putative alternative peptidoglycan biosynthesis pathway genes or any related enzyme. Finally, the mutants did not present an increased phosphoenolpyruvate concentration that might compete with fosfomycin for its binding to MurA. On the basis of these results, we describe a completely novel mechanism of antibiotic resistance based on mutations of genes encoding metabolic enzymes. IMPORTANCE Antibiotic resistance has been largely considered a specific bacterial response to an antibiotic challenge. Indeed, its study has been mainly concentrated on mechanisms that affect the antibiotics (mutations in transporters, efflux pumps, and antibiotic-modifying enzymes, or their regulators) or their targets (i.e., target mutations, protection, or bypass). Usually, antibiotic resistance-associated metabolic changes were considered a consequence (fitness costs) and not a cause of antibiotic resistance. Herein, we show that alterations in the central carbon bacterial metabolism can also be the cause of antibiotic resistance. In the study presented here, Stenotrophomonas maltophilia acquires fosfomycin resistance through the inactivation of glycolytic enzymes belonging to the Embden-Meyerhof-Parnas pathway. Besides resistance to fosfomycin, this inactivation also impairs the bacterial gluconeogenic pathway. Together with previous work showing that antibiotic resistance can be under metabolic control, our results provide evidence that antibiotic resistance is intertwined with the bacterial metabolism.

scholarly journals Development of a DNA Microarray for Enterococcal Species, Virulence, and Antibiotic Resistance Gene Determinations among Isolates from Poultry

2011 ◽  
Vol 77 (8) ◽  
pp. 2625-2633 ◽  
Author(s):  
J. Champagne ◽  
M. S. Diarra ◽  
H. Rempel ◽  
E. Topp ◽  
C. W. Greer ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Dna Microarray ◽  
Virulence Genes ◽  
Antibiotic Resistance Gene ◽  
Antibiotic Resistance Genes ◽  
Content Type ◽  
Virulence Potential ◽  
Enterococcal Species ◽  
Species Specific ◽  
Simultaneous Identification

ABSTRACTA DNA microarray (Enteroarray) was designed with probes targeting four species-specific taxonomic identifiers to discriminate among 18 different enterococcal species, while other probes were designed to identify 18 virulence factors and 174 antibiotic resistance genes. In total, 262 genes were utilized for rapid species identification of enterococcal isolates, while characterizing their virulence potential through the simultaneous identification of endogenous antibiotic resistance and virulence genes. Enterococcal isolates from broiler chicken farms were initially identified by using the API 20 Strep system, and the results were compared to those obtained with the taxonomic genesatpA,recA,pheS, andddlrepresented on our microarray. Among the 171 isolates studied, five different enterococcal species were identified by using the API 20 Strep system:Enterococcus faecium,E. faecalis,E. durans,E. gallinarum, andE. avium. The Enteroarray detected the same species as API 20 Strep, as well as two more:E. casseliflavusandE. hirae. Species comparisons resulted in 15% (27 isolates) disagreement between the two methods among the five API 20 Strep identifiable species and 24% (42 isolates) disagreement when considering the seven Enteroarray identified species. The species specificity of key antibiotic and virulence genes identified by the Enteroarray were consistent with the literature adding further robustness to the redundant taxonomic probe data. Sequencing of thecpn60gene further confirmed the complete accuracy of the microarray results. The new Enteroarray should prove to be a useful tool to accurately genotype strains of enterococci and assess their virulence potential.

scholarly journals Plasmid with Colistin Resistance Genemcr-1in Extended-Spectrum-β-Lactamase-Producing Escherichia coli Strains Isolated from Pig Slurry in Estonia

2016 ◽  
Vol 60 (11) ◽  
pp. 6933-6936 ◽  
Author(s):  
Age Brauer ◽  
Kaidi Telling ◽  
Mailis Laht ◽  
Piret Kalmus ◽  
Irja Lutsar ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Resistance Gene ◽  
Antibiotic Resistance Gene ◽  
Dna Transfer ◽  
The Other ◽  
Pig Slurry ◽  
Colistin Resistance ◽  
Content Type ◽  
Slurry Sample

ABSTRACTA plasmid carrying the colistin resistance genemcr-1was isolated from a pig slurry sample in Estonia. The gene was present on a 33,311-bp plasmid of the IncX4 group.mcr-1is the only antibiotic resistance gene on the plasmid, with the other genes mainly coding for proteins involved in conjugative DNA transfer (taxA,taxB,taxC,trbM, and thepilXoperon). The plasmid pESTMCR was present in three phylogenetically very differentEscherichia colistrains, suggesting that it has high potential for horizontal transfer.

scholarly journals Housefly Larva Vermicomposting Efficiently Attenuates Antibiotic Resistance Genes in Swine Manure, with Concomitant Bacterial Population Changes

2015 ◽  
Vol 81 (22) ◽  
pp. 7668-7679 ◽  
Author(s):  
Hang Wang ◽  
Hongyi Li ◽  
Jack A. Gilbert ◽  
Haibo Li ◽  
Longhua Wu ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Waste Management ◽  
Resistance Genes ◽  
Management Practice ◽  
Antibiotic Resistance Gene ◽  
Tetracycline Resistance ◽  
Feed Additives ◽  
Content Type ◽  
Genes Encoding ◽  
Waste Management Practice

ABSTRACTManure from swine treated with antimicrobials as feed additives is a major source for the expansion of the antibiotic resistance gene (ARG) reservoir in the environment. Vermicomposting via housefly larvae (Musca domestica) can be efficiently used to treat manure and regenerate biofertilizer, but few studies have investigated its effect on ARG attenuation. Here, we tracked the abundances of 9 ARGs and the composition and structure of the bacterial communities in manure samples across 6 days of full-scale manure vermicomposting. On day 6, the abundances of genes encoding tetracycline resistance [tet(M),tet(O),tet(Q), andtet(W)] were reduced (P< 0.05), while those of genes encoding sulfonamide resistance (sul1andsul2) were increased (P< 0.05) when normalized to 16S rRNA. The abundances of tetracycline resistance genes were correlated (P< 0.05) with the changing concentrations of tetracyclines in the manure. The overall diversity and richness of the bacteria significantly decreased during vermicomposting, accompanied by a 100 times increase in the relative abundance ofFlavobacteriaceaespp. Variations in the abundances of ARGs were correlated with the changing microbial community structure and the relative abundances of the familyRuminococcaceae, classBacilli, or phylumProteobacteria. Vermicomposting, as a waste management practice, can reduce the overall abundance of ARGs. More research is warranted to assess the use of this waste management practice as a measure to attenuate the dissemination of antimicrobial residues and ARGs from livestock production before vermicompost can be safely used as biofertilizer in agroecosystems.

scholarly journals Complete Genome Sequence of Escherichia coli Antibiotic Resistance Isolate Bank Number 0346

2021 ◽  
Vol 10 (23) ◽  
Author(s):  
Keira L. Stuart ◽  
Darrell O. Bayles ◽  
Sarah M. Shore ◽  
Tracy L. Nicholson
Keyword(s):  
Public Health ◽  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Genome Sequence ◽  
Complete Genome Sequence ◽  
Horizontal Transfer ◽  
Complete Genome ◽  
Content Type ◽  
Health Concerns

Plasmid-mediated polymyxin resistance encoded by mcr-1 has increased public health concerns due to the potential for rapid horizontal transfer. Here, we report the complete genome sequence of colistin-resistant Escherichia coli Antibiotic Resistance Isolate Bank number 0346, harboring a plasmid-borne mcr-1 gene.

scholarly journals ARG-ANNOT, a New Bioinformatic Tool To Discover Antibiotic Resistance Genes in Bacterial Genomes

2013 ◽  
Vol 58 (1) ◽  
pp. 212-220 ◽  
Author(s):  
Sushim Kumar Gupta ◽  
Babu Roshan Padmanabhan ◽  
Seydina M. Diene ◽  
Rafael Lopez-Rojas ◽  
Marie Kempf ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Gene Sequence ◽  
Antibiotic Resistance Gene ◽  
Antibiotic Resistance Genes ◽  
Genetic Determinants ◽  
Bacterial Genomes ◽  
Content Type ◽  
A Genome ◽  
Bioinformatic Tool

ABSTRACTARG-ANNOT (Antibiotic Resistance Gene-ANNOTation) is a new bioinformatic tool that was created to detect existing and putative new antibiotic resistance (AR) genes in bacterial genomes. ARG-ANNOT uses a local BLAST program in Bio-Edit software that allows the user to analyze sequences without a Web interface. All AR genetic determinants were collected from published works and online resources; nucleotide and protein sequences were retrieved from the NCBI GenBank database. After building a database that includes 1,689 antibiotic resistance genes, the software was tested in a blind manner using 100 random sequences selected from the database to verify that the sensitivity and specificity were at 100% even when partial sequences were queried. Notably, BLAST analysis results obtained using thermtFgene sequence (a new aminoglycoside-modifying enzyme gene sequence that is not included in the database) as a query revealed that the tool was able to link this sequence to short sequences (17 to 40 bp) found in other genes of thermtfamily with significant E values. Finally, the analysis of 178Acinetobacter baumanniiand 20Staphylococcus aureusgenomes allowed the detection of a significantly higher number of AR genes than the Resfinder gene analyzer and 11 point mutations in target genes known to be associated with AR. The average time for the analysis of a genome was 3.35 ± 0.13 min. We have created a concise database for BLAST using a Bio-Edit interface that can detect AR genetic determinants in bacterial genomes and can rapidly and easily discover putative new AR genetic determinants.

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