Transduction of bacteriophage resistance in Streptococcus pyogenes with coinfection lysates of the virulent phage A25ts1-2 and a temperate bacteriophage 2511

1984 ◽  
Vol 30 (11) ◽  
pp. 1309-1314
Author(s):  
James G. Stuart ◽  
Rene Rogel ◽  
Tom Morton
Keyword(s):  
Antibiotic Resistance ◽  
Streptococcus Pyogenes ◽  
Antibiotic Resistance Genes ◽  
Clinical Strain ◽  
Phage Resistance ◽  
Lytic Phage ◽  
The Novel ◽  
Temperate Bacteriophage ◽  
Chromosomal Dna ◽  
Viral Dna

Two new mutants of Streptococcus pyogenes designated KS107 and KS136 were shown to allow adsorption of a virulent bacteriophage KV1, but replication of viral DNA did not occur in these mutants. The novel observation was made that coinfection of donor ceils with phages A25ts1-2 and 2511 enhanced transduction of phage resistance and of antibiotic resistance. Mixed donor lysates did not give enhanced transduction. Antiserum to A25ts1-2 blocked transduction, suggesting that the presence of 2511 promoted packaging of chromosomal DNA into A25ts1-2 particles. Phage 2511 was isolated by induction of a clinical strain, but it behaved as a lytic phage in laboratory strains. Phage KV1 was isolated from sewage and was found to be closely related to A25ts1-2, whereas 2511 was clearly different. The phage resistance gene of KS107 was not linked to any of three antibiotic resistance genes by cotransduction tests.

scholarly journals Colocation of the Multiresistance Gene cfr and the Fosfomycin Resistance Gene fosD on a Novel Plasmid in Staphylococcus arlettae from a Chicken Farm

2017 ◽  
Vol 61 (12) ◽  
Author(s):  
Bi-Hui Liu ◽  
Chang-Wei Lei ◽  
An-Yun Zhang ◽  
Yun Pan ◽  
Ling-Han Kong ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Gene ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
The Novel ◽  
Content Type ◽  
Chicken Farm ◽  
Fosfomycin Resistance ◽  
First Time

ABSTRACT The novel 63,558-bp plasmid pSA-01, which harbors nine antibiotic resistance genes, including cfr, erm(C), tet(L), erm(T), aadD, fosD, fexB, aacA-aphD, and erm(B), was characterized in Staphylococcus arlettae strain SA-01, isolated from a chicken farm in China. The colocation of cfr and fosD genes was detected for the first time in an S. arlettae plasmid. The detection of two IS431-mediated circular forms containing resistance genes in SA-01 suggested that IS431 may facilitate dissemination of antibiotic resistance genes.

scholarly journals Molecular Evolution in a Multidrug-Resistant Lineage of Streptococcus pneumoniae: Emergence of Strains Belonging to the Serotype 6B Icelandic Clone That Lost Antibiotic Resistance Traits

2000 ◽  
Vol 38 (4) ◽  
pp. 1375-1381 ◽  
Author(s):  
Sigurdur E. Vilhelmsson ◽  
Alexander Tomasz ◽  
Karl G. Kristinsson
Keyword(s):  
Antibiotic Resistance ◽  
Streptococcus Pneumoniae ◽  
Pneumococcal Disease ◽  
Antibiotic Resistance Genes ◽  
Multidrug Resistant ◽  
Chromosomal Dna ◽  
Surveillance Period ◽  
Complete Collection ◽  
Predominant Type ◽  
Resistance To Penicillin

Since their first detection in 1988, penicillin-resistantStreptococcus pneumoniae isolates have rapidly spread in Iceland to account for close to 20% of all pneumococcal disease in that country by 1993. The major component (70%) of the resistant pneumococci identified from 1989 to 1992 was the progeny of a single multidrug-resistant clone (Icelandic clone) with a homogeneous chromosomal macrorestriction profile and identical multilocus enzyme type expressing serotype 6B and resistance to penicillin, tetracycline, chloramphenicol, erythromycin, and trimethoprim-sulfamethoxazole. The rest of the non-penicillin-susceptible isolates included bacteria with serotype 6A and serogroups 19 and 23. The unique geographic and epidemiological setting and the availability of a complete collection of all non-penicillin-susceptible isolates of S. pneumoniaein Iceland prompted us to carry out a molecular epidemiological study to monitor the fate of the Icelandic clone between 1989 and 1996; in addition, we wished to extend the characterization to representative groups of all non-penicillin-susceptible serotype 6B pneumococci which showed variations in antibiotype and which were recovered in Iceland between late 1989 and the end of 1996. Also included in the study were non-penicillin-susceptible isolates of serogroup 23. Pulsed-field gel electrophoresis of SmaI-restricted chromosomal DNA and Southern hybridization with the lytA DNA probe and probes specific for antibiotic resistance genes were used to characterize pneumococcal isolates. The results show that (i) the Icelandic clone remained the predominant type among penicillin-resistant S. pneumoniae through 1996; (ii) the emergence of variants of the Icelandic clone which had lost one or more of the antibiotic resistance phenotypes and/or resistant genes, singly or in combination, was documented during the surveillance period; and (iii) isolates belonging to the internationally spread multidrug-resistant serotype 23F clone were present in the Icelandic collection since late 1989 but did not increase in number during the subsequent years.

scholarly journals Novel Variants of AbaR Resistance Islands with a Common Backbone in Acinetobacter baumannii Isolates of European Clone II

2012 ◽  
Vol 56 (4) ◽  
pp. 1969-1973 ◽  
Author(s):  
Vaida Šeputienė ◽  
Justas Povilonis ◽  
Edita Sužiedėlienė
Keyword(s):  
Antibiotic Resistance ◽  
Acinetobacter Baumannii ◽  
Stress Protein ◽  
Antibiotic Resistance Genes ◽  
Structural Similarity ◽  
Chromosomal Dna ◽  
Class 1 Integron ◽  
Content Type ◽  
Novel Variants ◽  
Class 1

ABSTRACTIn this study, the genetic organization of three novel genomic antibiotic resistance islands (AbaRs) inAcinetobacter baumanniiisolates belonging to group of European clone II (EC II)comMintegrated sequences of 18-, 21-, and 23-kb resistance islands were determined. These resistance islands carry the backbone of AbaR-type transposon structures, which are composed of the transposition module coding for potential transposition proteins and other genes coding for the intact universal stress protein (uspA), sulfate permease (sul), and proteins of unknown function. The antibiotic resistance genesstrA,strB,tetB, andtetRand insertion sequence CR2 element were found to be inserted into the AbaR transposons. GenBank homology searches indicated that they are closely related to the AbaR sequences found integrated incomMin strains of EC II (A. baumanniistrains 1656-2 and TCDC-AB0715) and AbaR4 integrated in another location ofA. baumanniiAB0057 (EC I). All of the AbaRs showed structural similarity to the previously described AbaR4 island and share a 12,008-bp backbone. AbaRs contain Tn1213, Tn2006, and the multiple fragments which could be derived from transposons Tn3, Tn10, Tn21, Tn1000, Tn5393, and Tn6020, the insertion sequences IS26, ISAba1, ISAba14, and ISCR2, and the class 1 integron. Moreover, chromosomal DNA was inserted into distinct regions of the AbaR backbone. Sequence analysis suggested that the AbaR-type transposons have evolved through insertions, deletions, and homologous recombination. AbaR islands, sharing the core structure similar to AbaR4, appeared to be distributed in isolates of EC I and EC II via integration into distinct genomic sites, i.e.,phoandcomM, respectively.

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 ramR Deletion in an Enterobacter hormaechei Isolate as a Consequence of Therapeutic Failure of Key Antibiotics in a Long-Term Hospitalized Patient

2020 ◽  
Vol 64 (10) ◽  
Author(s):  
François Gravey ◽  
Vincent Cattoir ◽  
Frédéric Ethuin ◽  
Laetitia Fabre ◽  
Racha Beyrouthy ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Antibiotic Resistance Genes ◽  
Cross Resistance ◽  
Clinical Strain ◽  
Nucleotide Polymorphisms ◽  
Content Type ◽  
Expression Of Genes ◽  
Reverse Transcription Quantitative Pcr ◽  
Enterobacter Hormaechei ◽  
Hospitalized Patient

ABSTRACT Genome changes are central to the adaptation of bacteria, especially under antibiotic pressure. The aim of this study was to report phenotypic and genomic adaptations undergone by an Enterobacter hormaechei clinical strain that became highly resistant to key antimicrobials during a 4-month period in a patient hospitalized in an intensive care unit (ICU). All six clinical E. hormaechei strains isolated in one ICU-hospitalized patient have been studied. MICs regarding 17 antimicrobial molecules have been measured. Single nucleotide polymorphisms (SNPs) were determined on the sequenced genomes. The expression of genes involved in antibiotic resistance among Enterobacter cloacae complex strains were determined by reverse transcription-quantitative PCR (qRT-PCR). All the strains belonged to sequence type 66 and were distant by a maximum of nine SNPs. After 3 months of hospitalization, three strains presented a significant increase in MICs for ceftazidime, cefepime, temocillin, ertapenem, tigecycline, ciprofloxacin, and chloramphenicol. Those resistant strains did not acquire additional antibiotic resistance genes but harbored a 16-bp deletion in the ramR gene. This deletion led to upregulated expression of RamA, AcrA, AcrB, and TolC and downregulated expression of OmpF. The ΔramR mutant harbored the same phenotype as the resistant clinical strains regarding tigecycline, chloramphenicol, and ciprofloxacin. The increased expression of RamA due to partial deletion in the ramR gene led to a cross-resistance phenotype by an increase of antibiotic efflux through the AcrAB-TolC pump and a decrease of antibiotic permeability by porin OmpF. ramR appears to be an important adaptative trait for E. hormaechei strains.

Assessing the heterogeneity of in silico plasmid predictions based on whole-genome-sequenced clinical isolates

2017 ◽  
Vol 20 (3) ◽  
pp. 857-865 ◽  
Author(s):  
Cedric C Laczny ◽  
Valentina Galata ◽  
Achim Plum ◽  
Andreas E Posch ◽  
Andreas Keller
Keyword(s):  
Antibiotic Resistance ◽  
In Silico ◽  
Pathogenic Bacteria ◽  
Antibiotic Resistance Genes ◽  
Clinical Isolates ◽  
Whole Genome ◽  
Chromosomal Dna ◽  
Accurate Identification ◽  
Resistance Factors ◽  
The Individual

AbstractHigh-throughput next-generation shotgun sequencing of pathogenic bacteria is growing in clinical relevance, especially for chromosomal DNA-based taxonomic identification and for antibiotic resistance prediction. Genetic exchange is facilitated for extrachromosomal DNA, e.g. plasmid-borne antibiotic resistance genes. Consequently, accurate identification of plasmids from whole-genome sequencing (WGS) data remains one of the major challenges for sequencing-based precision medicine in infectious diseases. Here, we assess the heterogeneity of four state-of-the-art tools (cBar, PlasmidFinder, plasmidSPAdes and Recycler) for the in silico prediction of plasmid-derived sequences from WGS data. Heterogeneity, sensitivity and precision were evaluated by reference-independent and reference-dependent benchmarking using 846 Gram-negative clinical isolates. Interestingly, the majority of predicted sequences were tool-specific, resulting in a pronounced heterogeneity across tools for the reference-independent assessment. In the reference-dependent assessment, sensitivity and precision values were found to substantially vary between tools and across taxa, with cBar exhibiting the highest median sensitivity (87.45%) but a low median precision (27.05%). Furthermore, integrating the individual tools into an ensemble approach showed increased sensitivity (95.55%) while reducing the precision (25.62%). CBar and plasmidSPAdes exhibited the strongest concordance with respect to identified antibiotic resistance factors. Moreover, false-positive plasmid predictions typically contained only few antibiotic resistance factors. In conclusion, while high degrees of heterogeneity and variation in sensitivity and precision were observed across the different tools and taxa, existing tools are valuable for investigating the plasmid-borne resistome. Nevertheless, additional studies on representative clinical data sets will be necessary to translate in silico plasmid prediction approaches from research to clinical application.

scholarly journals Effects of the Newly Isolated T4-Like Phage on Transmission of Plasmid-Borne Antibiotic Resistance Genes via Generalized Transduction

Viruses ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 2070
Author(s):  
Junxuan Zhang ◽  
Xiaolu He ◽  
Shuqing Shen ◽  
Mengya Shi ◽  
Qin Zhou ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Insertion Site ◽  
Antibiotic Resistance Genes ◽  
Lytic Phage ◽  
Long Read ◽  
Biological Entities ◽  
Low Coverage ◽  
Generalized Transduction ◽  
Phage Population

Bacteriophages are the most abundant biological entities on earth and may play an important role in the transmission of antibiotic resistance genes (ARG) from host bacteria. Although the specialized transduction mediated by the temperate phage targeting a specific insertion site is widely explored, the carrying characteristics of “transducing particles” for different ARG subtypes in the process of generalized transduction remains largely unclear. Here, we isolated a new T4-like lytic phage targeting transconjugant Escherichia coli C600 that contained plasmid pHNAH67 (KX246266) and encoded 11 different ARG subtypes. We found that phage AH67C600_Q9 can misload plasmid-borne ARGs and package host DNA randomly. Moreover, for any specific ARG subtype, the carrying frequency was negatively correlated with the multiplicity of infection (MOI). Further, whole genome sequencing (WGS) identified that only 0.338% (4/1183) of the contigs of an entire purified phage population contained ARG sequences; these were floR, sul2, aph(4)-Ia, and fosA. The low coverage indicated that long-read sequencing methods are needed to explore the mechanism of ARG transmission during generalized transduction.

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 Determination the site of antibiotic resistance genes in Escherichia coli isolated From Urinary Tract Infection

2018 ◽  
Vol 3 (2) ◽  
pp. 6-12
Author(s):  
Karzan Abdulmuhsin Mohammad ◽  
Zirak F. Ahmed ◽  
Bayar A Mohammed ◽  
Rasti H Saeed
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Urinary Tract ◽  
Nalidixic Acid ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
Diffusion Method ◽  
Resistant Bacteria ◽  
Screening Methods ◽  
Chromosomal Dna

This study includes isolation of 25 isolates of Escherichia coli (E. coli  ) strain from urinary tract samples in a pregnant woman. Microbiological and biochemical tests were used to identify the resistant bacteria of this genus. Screening methods were used to determine bacterial isolates for their resistance to 10 antibiotics include: Amikacin (Ak), Amoxicillin (Ax), Ampicillin (Ap), Chloramphenicol (Cm), Ciprofloxacin (Cip), Erythromycin (Er), Nalidixic acid (Nal), Penicillin (Pen), Tetracycline (Tet) and Trimethoprim (Tm). The isolates E4, E9, E16, and E17 were resistant to all antibiotics used in the current  study using the disk diffusion method. In contrast, the resistance percentage for all antibiotics ranged between 28-96%. Sites of resistance genes and hemolysin production genes were characterized by tranformation techniques in the E4 and E16. The results showed that the antibiotic resistance genes of Amikacin, Erythromycin, Tetracyclin, and Trimethoprim were located on a plasmid, whereas Amoxicillin, Ampicillin, Chloramphenicol, Ciprofloxacin, Nalidixic acid and Penicillin were located on chromosomal DNA. The results also demonstrated an inability to produce alpha or beta-hemolysin indicating that the genes which are responsible for hemolysin production were also located on chromosomal DNA. 

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