Ten Enterobacteriaceae isolates collected in a Czech hospital carried blaKPC-positive plasmids of different sizes (∼30, ∼45, and ∼80 kb). Sequencing revealed three types of plasmids (A to C) with the Tn4401a transposon. Type A plasmids comprised an IncR backbone and a KPC-2-encoding multidrug resistance (MDR) region. Type B plasmids were derivatives of type A plasmids carrying an IncN3-like segment, while type C plasmids were IncP6 plasmids sharing the same KPC-2-encoding MDR region with type A and B plasmids.
- MeSH
- Anti-Bacterial Agents therapeutic use MeSH
- beta-Lactamases genetics metabolism MeSH
- Citrobacter freundii drug effects enzymology genetics isolation & purification MeSH
- Enterobacteriaceae Infections drug therapy epidemiology microbiology MeSH
- Escherichia coli drug effects enzymology genetics isolation & purification MeSH
- Gene Expression MeSH
- Isoenzymes genetics metabolism MeSH
- Carbapenems therapeutic use MeSH
- Klebsiella pneumoniae drug effects enzymology genetics isolation & purification MeSH
- Humans MeSH
- Microbial Sensitivity Tests MeSH
- Drug Resistance, Multiple, Bacterial genetics MeSH
- Morganella morganii drug effects enzymology genetics isolation & purification MeSH
- Hospitals MeSH
- Open Reading Frames MeSH
- Plasmids chemistry classification metabolism MeSH
- Base Sequence MeSH
- Sequence Analysis, DNA MeSH
- DNA Transposable Elements MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Czech Republic MeSH
OBJECTIVES: To describe the prevalence and diversity of IncX plasmids with antibiotic resistance genes in Enterobacteriaceae and to identify the most disseminated lineages of the plasmid family. METHODS: IncX plasmids were screened in 1894 Enterobacteriaceae isolates resistant to cefotaxime (2 mg/L) or with reduced susceptibility to ciprofloxacin (0.05 mg/L) obtained from various sources in five continents using PCR. IncX plasmid-harbouring isolates were identified using MALDI-TOF or biochemical tests, and screened for antibiotic resistance genes using PCR and sequencing; their clonality was determined by PFGE. Horizontal transfer of plasmids was tested using transformation and conjugation. IncX plasmids were characterized by S1-nuclease and PFGE, RFLP and hybridization. RESULTS: A total of 164 Escherichia coli isolates (8.7%, n = 1894) carried at least one IncX subgroup. Seven isolates harboured two distinct subgroups. IncX1 subgroup was found in 93 isolates, followed by IncX2 (35 isolates), IncX4 (28) and IncX3 (15). IncX4 plasmids were not transferred horizontally as single plasmids and therefore excluded from further analysis. The most disseminated lineages of IncX plasmids included IncX1 harbouring qnrS1 and blaTEM-1,-135 found in 36 E. coli from different sources in Europe and Australia and IncX2 carrying qnrS1 and tet(A) detected in nine E. coli from wildlife in Europe. IncX3 plasmids harboured predominantly blaSHV-12 and qnrS1 or qnrB7. CONCLUSIONS: IncX plasmids were widely distributed in E. coli from wildlife in Europe and were predominantly associated with fluoroquinolone resistance genes. Plasmids showing indistinguishable restriction profiles were identified in E. coli from different sources and countries suggesting wide dissemination of certain plasmid lineages.
- MeSH
- Genes, Bacterial * MeSH
- Drug Resistance, Bacterial * MeSH
- Transformation, Bacterial MeSH
- beta-Lactams pharmacology MeSH
- Global Health MeSH
- Animals, Wild MeSH
- Escherichia coli drug effects genetics isolation & purification MeSH
- Fluoroquinolones pharmacology MeSH
- Genetic Variation MeSH
- Genotype MeSH
- Escherichia coli Infections microbiology veterinary MeSH
- Conjugation, Genetic MeSH
- Humans MeSH
- Molecular Typing MeSH
- Plasmids analysis classification MeSH
- Polymerase Chain Reaction MeSH
- Gene Transfer, Horizontal MeSH
- Prevalence MeSH
- Electrophoresis, Gel, Pulsed-Field MeSH
- Sequence Analysis, DNA MeSH
- Cluster Analysis MeSH
- Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
This study concerned the occurrence of fecal bacteria with plasmid-mediated quinolone resistance (PMQR) genes in rooks (Corvus frugilegus, medium-sized corvid birds) wintering in continental Europe during winter 2010/2011. Samples of fresh rook feces were taken by cotton swabs at nine roosting places in eight European countries. Samples were transported to one laboratory and placed in buffered peptone water (BPW). The samples from BPW were enriched and subcultivated onto MacConkey agar (MCA) supplemented with ciprofloxacin (0.06 mg/L) to isolate fluoroquinolone-resistant bacteria. DNA was isolated from smears of bacterial colonies growing on MCA and tested by PCR for PMQR genes aac(6')-Ib, qepA, qnrA, qnrB, qnrC, qnrD, qnrS, and oqxAB. All the PCR products were further analyzed by sequencing. Ciprofloxacin-resistant bacteria were isolated from 37% (392 positive/1,073 examined) of samples. Frequencies of samples with ciprofloxacin-resistant isolates ranged significantly from 3% to 92% in different countries. The qnrS1 gene was found in 154 samples and qnrS2 in 2 samples. The gene aac(6')-Ib-cr was found in 16 samples. Thirteen samples were positive for qnrB genes in variants qnrB6 (one sample), qnrB18 (one), qnrB19 (one), qnrB29 (one), and qnrB49 (new variant) (one). Both the qnrD and oqxAB genes were detected in six samples. The genes qnrA, qnrC, and qepA were not found. Wintering omnivorous rooks in Europe were commonly colonized by bacteria supposedly Enterobacteriaceae with PMQR genes. Rooks may disseminate these epidemiologically important bacteria over long distances and pose a risk for environmental contamination.
- MeSH
- Anti-Bacterial Agents pharmacology MeSH
- Enterobacteriaceae genetics isolation & purification MeSH
- Enterobacteriaceae Infections epidemiology microbiology veterinary MeSH
- Feces microbiology MeSH
- Fluoroquinolones pharmacology MeSH
- Microbial Sensitivity Tests MeSH
- Bird Diseases epidemiology microbiology MeSH
- Plasmids classification genetics isolation & purification MeSH
- Polymerase Chain Reaction veterinary MeSH
- Protein Isoforms classification genetics isolation & purification MeSH
- Escherichia coli Proteins classification genetics isolation & purification MeSH
- Crows microbiology MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Europe MeSH
Complete sequences for two plasmids associated with two strains of "Candidatus Phytoplasma asteris" have been obtained. The plasmid named pPARG1 was found in Rehmannia glutinosa L. associated with phytoplasma classified in the 16Sr I-C subgroup. Plasmid pPABN1 was from phytoplasma associated with infected winter oilseed rape and classified in the 16Sr I-B subgroup. The plasmids pPARG1 (4371 nt) and pPABN1 (3529 nt) have high A+T content of about 75%, similar to that of phytoplasma genomes. Six and five open reading frames longer than 100 amino acids and organized on the same strand were recognized on pPARG1 and pPABN1, respectively. A segment about 1.6 kbp long sharing high sequence identity with the Onion yellows phytoplasma genome was found in pPABN1.
- MeSH
- Brassica napus microbiology MeSH
- Phylogeny MeSH
- Host-Pathogen Interactions MeSH
- Molecular Sequence Data MeSH
- Plant Diseases microbiology MeSH
- Open Reading Frames MeSH
- Phytoplasma genetics pathogenicity MeSH
- Plasmids classification genetics MeSH
- Rehmannia genetics microbiology MeSH
- Plants microbiology MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- MeSH
- Drug Resistance, Microbial MeSH
- Salmonella Phages genetics isolation & purification classification MeSH
- Humans MeSH
- Plasmids physiology isolation & purification classification MeSH
- Salmonella enteritidis classification drug effects virology MeSH
- Salmonella Infections microbiology MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
- MeSH
- DNA, Bacterial genetics classification metabolism MeSH
- Humans MeSH
- Plasmids genetics classification MeSH
- Deoxyribonucleases, Type II Site-Specific MeSH
- Salmonella enteritidis genetics classification pathology MeSH
- Salmonella Infections, Animal microbiology MeSH
- Bacterial Typing Techniques MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Animals MeSH
