scholarly journals Mutations of domain V in 23S ribosomal RNA of macrolide-resistant Mycoplasma gallisepticum isolates in Egypt

2016 ◽  
Vol 10 (08) ◽  
pp. 807-813 ◽  
Author(s):  
Ahmed M Ammar ◽  
Norhan K Abd El-Aziz ◽  
Ahlam A Gharib ◽  
Hanaa K Ahmed ◽  
Amira E Lameay
Keyword(s):  
Mycoplasma Gallisepticum ◽  
23S Rrna ◽  
Nucleotide Sequencing ◽  
Rrna Gene ◽  
Intrinsic Resistance ◽  
Respiratory Problems ◽  
E Coli ◽  
23S Rrna Gene ◽  
Transversion Mutation ◽  
Domain V

Introduction: Avian mycoplasmas impose a significant economic burden to the poultry industry. In recent years, macrolide-resistant Mycoplasma gallisepticum have occasionally been encountered in Egypt. Methodology: This study was designed to document the involvement of macrolide-resistant M. gallisepticum in respiratory organs of chickens suffering respiratory problems. Concurrently, an exhaustive molecular characterization of the intrinsic resistance of recovered isolates to macrolides was done. Results: Of 120 chickens showing respiratory problems, 14 (11.67%) M. gallisepticum were isolated and genetically identified; 8 of them were recovered from air sacs, 4 from lungs, and 2 from tracheas. Broth microdilution of all M. gallisepticum isolates showed various degrees of minimum inhibitory concentrations (MICs) against macrolides: erythromycin (0.25–32 µg/mL), tylosin (0.0625–4 µg/mL), and tiamulin (0.031–2 µg/mL). Nucleotide sequencing of domain V (peptidyl transferase region) of the 23S rRNA gene of macrolide-resistant M. gallisepticum isolates revealed transition mutations at positions 2068 and 2069 (corresponding to 2058 and 2059 in Escherichia coli numbering) in an isolate and at position 2067 (corresponding to 2057 in E. coli numbering) in three isolates as hot spots for macrolide resistance. Surprisingly, a transversion mutation at position 2621 (corresponding to 2611 in E. coli numbering) was reported in one of the recovered isolates as a first report. Conclusion: Generation of new mutations is evidence for persistence of M. gallisepticum despite macrolide treatment. Periodic surveys to monitor for the possible appearance of resistant strains are recommended.

scholarly journals Characterization and Molecular Analysis of Macrolide-Resistant Mycoplasma pneumoniae Clinical Isolates Obtained in Japan

2004 ◽  
Vol 48 (12) ◽  
pp. 4624-4630 ◽  
Author(s):  
Mayumi Matsuoka ◽  
Mitsuo Narita ◽  
Norio Okazaki ◽  
Hitomi Ohya ◽  
Tsutomu Yamazaki ◽  
...  
Keyword(s):  
Mycoplasma Pneumoniae ◽  
Ribosomal Proteins ◽  
Respiratory Infections ◽  
Point Mutations ◽  
23S Rrna ◽  
Nucleotide Sequencing ◽  
Rrna Gene ◽  
Pcr Products ◽  
23S Rrna Gene ◽  
Domain V

ABSTRACT In recent years, Mycoplasma pneumoniae strains that are clinically resistant to macrolide antibiotics have occasionally been encountered in Japan. Of 76 strains of M. pneumoniae isolated in three different areas in Japan during 2000 to 2003, 13 strains were erythromycin (ERY) resistant. Of these 13 strains, 12 were highly ERY resistant (MIC, ≥256 μg/ml) and 1 was weakly resistant (MIC, 8 μg/ml). Nucleotide sequencing of domains II and V of 23S rRNA and ribosomal proteins L4 and L22, which are associated with ERY resistance, showed that 10 strains had an A-to-G transition at position 2063 (corresponding to 2058 in Escherichia coli numbering), 1 strain showed A-to-C transversion at position 2063, 1 strain showed an A-to-G transition at position 2064, and the weakly ERY-resistant strain showed C-to-G transversion at position 2617 (corresponding to 2611 in E. coli numbering) of domain V. Domain II and ribosomal proteins L4 and L22 were not involved in the ERY resistance of these clinical M. pneumoniae strains. In addition, by using our established restriction fragment length polymorphism technique to detect point mutations of PCR products for domain V of the 23S rRNA gene of M. pneumoniae, we found that 23 (24%) of 94 PCR-positive oral samples taken from children with respiratory infections showed A2063G mutation. These results suggest that ERY-resistant M. pneumoniae infection is not unusual in Japan.

scholarly journals Drug Resistance Mechanisms ofMycoplasma pneumoniaeto Macrolide Antibiotics

2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
Author(s):  
Xijie Liu ◽  
Yue Jiang ◽  
Xiaogeng Chen ◽  
Jing Li ◽  
Dawei Shi ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Resistance Mechanisms ◽  
23S Rrna ◽  
Macrolide Antibiotics ◽  
Rrna Gene ◽  
Site Mutation ◽  
23S Rrna Gene ◽  
Resistant Strains ◽  
Domain V ◽  
The 16S Rrna Gene

Throat swabs from children with suspectedMycoplasma pneumoniae(M. pneumoniae) infection were cultured for the presence ofM. pneumoniaeand its species specificity using the 16S rRNA gene. Seventy-sixM. pneumoniaestrains isolated from 580 swabs showed that 70 were erythromycin resistant with minimum inhibitory concentrations (MIC) around 32–512 mg/L. FiftyM. pneumoniaestrains (46 resistant, 4 sensitive) were tested for sensitivity to tetracycline, ciprofloxacin, and gentamicin. Tetracycline and ciprofloxacin had some effect, and gentamicin had an effect on the majority ofM. pneumoniaestrains. Domains II and V of the 23S rRNA gene and the ribosomal protein L4 and L22 genes, both of which are considered to be associated with macrolide resistance, were sequenced and the sequences were compared with the corresponding sequences in M129 registered with NCBI and the FH strain. The 70 resistant strains all showed a 2063 or 2064 site mutation in domain V of the 23S rRNA but no mutations in domain II. Site mutations of L4 or L22 can be observed in either resistant or sensitive strains, although it is not known whether this is associated with drug resistance.

Identification of a Novel Mutation Affecting Domain V of the 23S rRNA Gene inHelicobacter pylori

Helicobacter ◽  
2004 ◽  
Vol 9 (5) ◽  
pp. 396-399 ◽  
Author(s):  
Sonia Toracchio ◽  
Gitana M. Aceto ◽  
Renato Mariani-Costantini ◽  
Pasquale Battista ◽  
Leonardo Marzio
Keyword(s):  
Novel Mutation ◽  
23S Rrna ◽  
Rrna Gene ◽  
23S Rrna Gene ◽  
Domain V

scholarly journals Macrolide Resistance in Campylobacter jejuni and Campylobacter coli: Molecular Mechanism and Stability of the Resistance Phenotype

2005 ◽  
Vol 49 (7) ◽  
pp. 2753-2759 ◽  
Author(s):  
Amera Gibreel ◽  
Veronica N. Kos ◽  
Monika Keelan ◽  
Cathy A. Trieber ◽  
Simon Levesque ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Target Gene ◽  
Macrolide Resistance ◽  
23S Rrna ◽  
Rrna Gene ◽  
Campylobacter Coli ◽  
Resistance Level ◽  
Resistance Phenotype ◽  
E Coli ◽  
23S Rrna Gene

ABSTRACT A collection of 23 macrolide-resistant Campylobacter isolates from different geographic areas was investigated to determine the mechanism and stability of macrolide resistance. The isolates were identified as Campylobacter jejuni or Campylobacter coli based on the results of the hippurate biochemical test in addition to five PCR-based genotypic methods. Three point mutations at two positions within the peptidyl transferase region in domain V of the 23S rRNA gene were identified. About 78% of the resistant isolates exhibited an A→G transition at Escherichia coli equivalent base 2059 of the 23S rRNA gene. The isolates possessing this mutation showed a wide range of erythromycin and clarithromycin MICs. Thus, this mutation may incur a greater probability of treatment failure in populations infected by resistant Campylobacter isolates. Another macrolide-associated mutation (A→C transversion), at E. coli equivalent base 2058, was detected in about 13% of the isolates. An A→G transition at a position cognate with E. coli 23S rRNA base 2058, which is homologous to the A2142G mutation commonly described in Helicobacter pylori, was also identified in one of the C. jejuni isolates examined. In the majority of C. jejuni isolates, the mutations in the 23S rRNA gene were homozygous except in two cases where the mutation was found in two of the three copies of the target gene. Natural transformation demonstrated the transfer of the macrolide resistance phenotype from a resistant Campylobacter isolate to a susceptible Campylobacter isolate. Growth rates of the resulting transformants containing A-2058→C or A-2059→G mutations were similar to that of the parental isolate. The erythromycin resistance of six of seven representative isolates was found to be stable after successive subculturing in the absence of erythromycin selection pressure regardless of the resistance level, the position of the mutation, or the number of the mutated copies of the target gene. One C. jejuni isolate showing an A-2058→G mutation, however, reverted to erythromycin and clarithromycin susceptibility after 55 subcultures on erythromycin-free medium. Investigation of ribosomal proteins L4 and L22 by sequence analysis in five representative isolates of C. jejuni and C. coli demonstrated no significant macrolide resistance-associated alterations in either the L4 or the L22 protein that might explain either macrolide resistance or enhancement of the resistance level.

scholarly journals Mutations in 23S rRNA gene associated with decreased susceptibility to tiamulin and valnemulin in Mycoplasma gallisepticum

2010 ◽  
pp. no-no ◽  
Author(s):  
Bei-Bei Li ◽  
Jian-Zhong Shen ◽  
Xing-Yuan Cao ◽  
Yang Wang ◽  
Lei Dai ◽  
...  
Keyword(s):  
Mycoplasma Gallisepticum ◽  
23S Rrna ◽  
Rrna Gene ◽  
23S Rrna Gene

scholarly journals Emergence of Macrolide-Resistant Mycoplasma pneumoniae with a 23S rRNA Gene Mutation

2005 ◽  
Vol 49 (6) ◽  
pp. 2302-2306 ◽  
Author(s):  
Miyuki Morozumi ◽  
Keiko Hasegawa ◽  
Reiko Kobayashi ◽  
Nagako Inoue ◽  
Satoshi Iwata ◽  
...  
Keyword(s):  
Mycoplasma Pneumoniae ◽  
Acute Bronchitis ◽  
23S Rrna ◽  
Rrna Gene ◽  
Microdilution Method ◽  
23S Rrna Gene ◽  
Pediatric Outpatients ◽  
Tract Infections ◽  
Domain V ◽  
Epidemiol Infect

ABSTRACT A total of 195 Mycoplasma pneumoniae strains were isolated from 2,462 clinical specimens collected between April 2002 and March 2004 from pediatric outpatients with respiratory tract infections. Susceptibilities to six macrolide antibiotics (ML), telithromycin, minocycline, levofloxacin, and sitafloxacin were determined by the microdilution method using PPLO broth. A total of 183 M. pneumoniae isolates were susceptible to all agents and had excellent MIC90s in the following order: 0.00195 μg/ml for azithromycin and telithromycin, 0.0078 μg/ml for clarithromycin, 0.0156 μg/ml for erythromycin, 0.0625 μg/ml for sitafloxacin, 0.5 μg/ml for minocycline, and 1 μg/ml for levofloxacin. Notably, 12 ML-resistant M. pneumoniae strains were isolated from patients with pneumonia (10 strains) or acute bronchitis (2 strains). These strains showed resistance to ML with MICs of ≥1 μg/ml, except to rokitamycin. Transition mutations of A2063G or A2064G, which correspond to A2058 and A2059 in Escherichia coli, in domain V on the 23S rRNA gene in 11 ML-resistant strains were identified. By pulsed-field gel electrophoresis typing, these strains were classified into groups I and Vb, as described previously (A. Cousin-Allery, A. Charron, B. D. Barbeyrac, G. Fremy, J. S. Jensen, H. Renaudin, and C. Bebear, Epidemiol. Infect. 124:103-111, 2000). These findings suggest that excessive usage of MLs acts as a trigger to select mutations on the corresponding 23S rRNA gene with the resultant occurrence of ML-resistant M. pneumoniae. Monitoring ML susceptibilities for M. pneumoniae is necessary in the future.

Novel Genotypes in Helicobacter pylori Involving Domain V of the 23S rRNA Gene

Helicobacter ◽  
2007 ◽  
Vol 12 (5) ◽  
pp. 505-509 ◽  
Author(s):  
Leonardo Garrido ◽  
Hector Toledo
Keyword(s):  
Helicobacter Pylori ◽  
23S Rrna ◽  
Rrna Gene ◽  
23S Rrna Gene ◽  
Domain V

scholarly journals Diversity of Domain V of 23S rRNA Gene Sequence in Different Enterococcus Species

2000 ◽  
Vol 38 (11) ◽  
pp. 3991-3993 ◽  
Author(s):  
Sotirios Tsiodras ◽  
Howard S. Gold ◽  
Eoin P. G. Coakley ◽  
Christine Wennersten ◽  
Robert C. Moellering ◽  
...  
Keyword(s):  
16S Rrna Gene Sequencing ◽  
23S Rrna ◽  
Rrna Gene ◽  
Dna Encoding ◽  
Enterococcus Casseliflavus ◽  
23S Rrna Gene ◽  
Rrna Gene Sequencing ◽  
Biochemical Testing ◽  
Enterococcus Gallinarum ◽  
Domain V

The highly conserved central loop of domain V of 23S RNA (nucleotides 2042 to 2628; Escherichia coli numbering) is implicated in peptidyltransferase activity and represents one of the target sites for macrolide, lincosamide, and streptogramin B antibiotics. DNA encoding domain V (590 bp) of several species ofEnterococcus was amplified by PCR. Twenty enterococcal isolates were tested, including Enterococcus faecium (six isolates), Enterococcus faecalis, Enterococcus avium, Enterococcus durans, Enterococcus gallinarum, Enterococcus casseliflavus (two isolates of each), and Enterococcus raffinosus, Enterococcus mundtii, Enterococcus malodoratus, andEnterococcus hirae (one isolate of each). For all isolates, species identification by biochemical testing was corroborated by 16S rRNA gene sequencing. The sequence of domain V of the 23S rRNA gene from E. faecium and E. faecalis differed from those of all other enterococci. The domain V sequences of E. durans and E. hirae were identical. This was also true for E. gallinarum and E. casseliflavus. E. avium differed from E. casseliflavus by 23 bases, from E. durans by 16 bases, and from E. malodoratus by 2 bases. E. avium differed fromE. raffinosus by one base. Despite the fact that domain V is considered to be highly conserved, substantial differences were identified between several enterococcal species.

scholarly journals Clinical-Use-Associated Decrease in Susceptibility of Vancomycin-Resistant Enterococcus faecium to Linezolid: a Comparison with Quinupristin-Dalfopristin

2004 ◽  
Vol 48 (9) ◽  
pp. 3583-3585 ◽  
Author(s):  
Issam I. Raad ◽  
Hend A. Hanna ◽  
Ray Y. Hachem ◽  
Tanya Dvorak ◽  
Rebecca B. Arbuckle ◽  
...  
Keyword(s):  
Gene Mutation ◽  
Enterococcus Faecium ◽  
23S Rrna ◽  
Rrna Gene ◽  
Clinical Use ◽  
Vancomycin Resistant Enterococcus ◽  
23S Rrna Gene ◽  
Vancomycin Resistant ◽  
Domain V

ABSTRACT The susceptibility of 135 vancomycin-resistant Enterococcus faecium bacteremic isolates to linezolid and quinupristin-dalfopristin was determined. All were susceptible to linezolid, while 88% were susceptible to quinupristin-dalfopristin prior to the clinical use of the drugs at our hospital. More than 6 months after their clinical use, a decrease in susceptibility was noted for only linezolid at 83%. This was related in part to a single G2576U gene mutation in domain V of the 23S rRNA gene.

scholarly journals Restriction Profiling of 23S Microheterogenic Ribosomal Repeats for Detection and Characterizing ofE. coliand Their Clonal, Pathogenic, and Phylogroups

2015 ◽  
Vol 2015 ◽  
pp. 1-11
Author(s):  
Parvathi Jayasree Rajagopalan Nair ◽  
Sunita Singh
Keyword(s):  
Cost Effective ◽  
23S Rrna ◽  
Rrna Gene ◽  
Type I ◽  
Microbial Identification ◽  
E Coli ◽  
Type Iv ◽  
Cost Effective Approach ◽  
23S Rrna Gene ◽  
K 12

Correlating ribosomal microheterogenicity with unique restriction profiles can prove to be an efficacious and cost-effective approach compared with sequencing for microbial identification. An attempt to peruse restriction profiling of 23S ribosomal assemblage was ventured; digestion patterns withBfaI discriminatedE. colifrom its colony morphovars, whileHaeIII profiles assisted in establishing distinct clonal groups. Among the gene pool of 399 ribosomal sequences extrapolated from 57E. coligenomes, varying degree of predominance (I > III > IV > II) ofHaeIII pattern was observed. This was also corroborated in samples collected from clinical, commensal, and environmental origin. K-12 and its descendants showed type I pattern whereasE. coli-B and its descendants exhibited type IV, both of these patterns being exclusively present inE. coli. A near-possible association between phylogroups andHaeIII profiles with presumable correlation between the clonal groups and different pathovars was established. The generic nature, conservation, and barcode gap of 23S rRNA gene make it an ideal choice and substitute to 16S rRNA gene, the most preferred region for molecular diagnostics in bacteria.

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