Molecular Typing and Macrolide Resistance Analyses ofTreponema pallidumin Heterosexuals and Men Who Have Sex with Men in Japan, 2017

ABSTRACTIn recent years, syphilis notifications have increased dramatically in Japan. We carried out molecular typing and macrolide resistance analyses ofTreponema pallidumsubsp.pallidumsamples collected from patients at four clinics and a hospital in Tokyo and Osaka prefectures in 2017. The macrolide resistant strain type 14d/f (SS14-like clade) was found in significantly more cases of syphilis among heterosexuals than in those among men who have sex with men (MSM); i.e., 79% (31/39) of the strains from heterosexuals were 14d/f compared to 37% (7/19) of those from MSM (odds ratio [OR], 6.6; 95% confidence interval [CI], 1.7 to 26.7;P = 0.002). In addition, 83% (50/60) of the strains were identified as macrolide resistant with an A2058G mutation in the 23S rRNA gene; 90% (35/39) of the strains from heterosexuals were macrolide resistant compared to 58% (11/19) of those from MSM. The odds of having the resistant mutation were considerably higher in the former (OR, 6.4; 95% CI, 1.3 to 33.5;P = 0.02). Heterosexual women and heterosexual men showed similar distributions, and the association remained the same when restricted to men. The strain type distribution and the prevalence of macrolide resistance differed substantially between syphilis strains from heterosexual cases and from MSM cases, suggesting distinct epidemiologic profiles for the two communities and providing important insight into the dynamics of syphilis in Japan.

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Molecular typing and macrolide resistance analyses ofTreponema pallidumin heterosexuals and men who have sex with men communities in Japan, 2017

10.1101/375485 ◽

2018 ◽

Author(s):

Mizue Kanai ◽

Yuzo Arima ◽

Shingo Nishiki ◽

Ken Shimuta ◽

Ichiro Itoda ◽

...

Keyword(s):

Molecular Typing ◽

Treponema Pallidum ◽

Macrolide Resistance ◽

23S Rrna ◽

Rrna Gene ◽

23S Rrna Gene ◽

Strain Type ◽

Sex With Men ◽

Insight Into ◽

Resistant Mutation

AbstractIn recent years, syphilis notifications have increased dramatically in Japan. We performed molecular typing and analyzed macrolide resistance ofTreponema pallidumsamples collected from four clinics and a hospital in Tokyo and Osaka prefectures in 2017.Macrolide resistant strain type 14d/f was found significantly more in heterosexual syphilis cases compared with those strains identified in men who have sex with men (MSM) syphilis cases. The proportion of 14d/f among heterosexuals was 79% (31/39) compared to 37% (7/19) among MSM [OR, 6.6 (95% CI, 1.7 to 26.7); P=0.002]. 83% (50/60) of the strains were identified as macrolide resistant with an A2058G mutation at the 23S rRNA gene. 90% (35/39) of the heterosexual strains were macrolide resistant, relative to 58% (11/19) of MSM strains; the odds of having the resistant mutation was considerably higher in heterosexuals compared with MSM [OR, 6.4 (95% CI, 1.3 to 33.5); P=0.02]. Heterosexual females and males showed similar distributions, and the results remained the same when restricted to men.The strain type distribution and frequency of macrolide resistance differed substantially between heterosexual and MSM syphilis samples, suggesting distinct epidemiologic profiles for the two communities and insight into syphilis transmission dynamics in Japan.

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Evidence for Multidrug Resistance in NonpathogenicMycoplasmaSpecies Isolated from South African Poultry

Applied and Environmental Microbiology ◽

10.1128/aem.01660-18 ◽

2018 ◽

Vol 84 (21) ◽

Cited By ~ 6

Author(s):

Amanda Beylefeld ◽

Pamela Wambulawaye ◽

Dauda Garba Bwala ◽

Johannes Jacobus Gouws ◽

Obed Mooki Lukhele ◽

...

Keyword(s):

Multidrug Resistance ◽

South African ◽

Macrolide Resistance ◽

Quinolone Resistance ◽

23S Rrna ◽

Clear Correlation ◽

Rrna Gene ◽

Poultry Production ◽

Content Type ◽

23S Rrna Gene

ABSTRACTOne hundred seventy-eight mycoplasma strains isolated from South African poultry flocks between 2003 and 2015 were identified by full-genome sequencing and phylogenetic analysis of the 16S rRNA gene and were classified as follows:Mycoplasma gallisepticum(25%),M. gallinarum(25%),M. gallinaceum, (23%),M. pullorum(14%),M. synoviae(10%), andM. iners(3%), as well as oneAcheoplasma laidlawiistrain (1%). MIC testing was performed on the axenic samples, and numerous strains of each species were resistant to either chlortetracycline or tylosin or both, with variable sensitivity to enrofloxacin. The strains of all species tested remained sensitive to tiamulin, except for oneM. gallinaceumsample that demonstrated intermediate sensitivity. The mutation of A to G at position 2059 (A2059G) in the 23S rRNA gene, which is associated with macrolide resistance, was found in the South AfricanM. gallisepticumandM. synoviaestrains, as well as a clear correlation between macrolide resistance inM. gallinarumandM. gallinaceumand mutations G354A and G748A in the L4 ribosomal protein and 23S rRNA gene, respectively. No correlation between resistance and point mutations in the genes studied could be found forM. pullorum. Only a few strains were resistant to enrofloxacin, apart from oneM. synoviaestrain with point mutation D420N, which has been associated with quinolone resistance, and no other known markers for quinolone resistance were found in this study. Proportionally more antimicrobial-resistant strains were detected inM. gallinaceum,M. gallinarum, andM. pullorumthan inM. gallisepticumandM. synoviae. Of concern, threeM. gallinaceumstrains showed multidrug resistance to chlortetracycline, tylosin, and oxytetracycline.IMPORTANCENonpathogenic poultryMycoplasmaspecies are often overlooked due to their lesser impact on poultry health and production compared to the OIE-listed pathogenic strainsM. gallisepticumandM. synoviae. The use of antimicrobials as in-feed growth promoters and for the control of mycoplasmosis is common in poultry production across the world. Here, we provide evidence that certain nonpathogenicMycoplasmaspecies are acquiring multidrug resistance traits. This would have significant implications if these species, for which no vaccines are applied, are able to transfer their antibiotic resistance genes to other mycoplasmas and bacteria that may enter the human food chain.

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Mutational and Transcriptomic Changes Involved in the Development of Macrolide Resistance in Campylobacter jejuni

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01927-12 ◽

2012 ◽

Vol 57 (3) ◽

pp. 1369-1378 ◽

Cited By ~ 26

Author(s):

Haihong Hao ◽

Zonghui Yuan ◽

Zhangqi Shen ◽

Jing Han ◽

Orhan Sahin ◽

...

Keyword(s):

Campylobacter Jejuni ◽

Dna Microarray ◽

Ribosomal Proteins ◽

Macrolide Resistance ◽

23S Rrna ◽

Rrna Gene ◽

Content Type ◽

23S Rrna Gene ◽

Resistant Mutants ◽

Antibiotic Efflux

ABSTRACTMacrolide antibiotics are important for clinical treatment of infections caused byCampylobacter jejuni. Development of resistance to this class of antibiotics inCampylobacteris a complex process, and the dynamic molecular changes involved in this process remain poorly defined. Multiple lineages of macrolide-resistant mutants were selected by stepwise exposure ofC. jejunito escalating doses of erythromycin or tylosin. Mutations in target genes were determined by DNA sequencing, and the dynamic changes in the expression of antibiotic efflux transporters and the transcriptome ofC. jejuniwere examined by real-time reverse transcription-PCR, immunoblotting, and DNA microarray analysis. Multiple types of mutations in ribosomal proteins L4 and L22 occurred early during stepwise selection. On the contrary, the mutations in the 23S rRNA gene, mediating high resistance to macrolides, were observed only in the late-stage mutants. Upregulation of antibiotic efflux genes was observed in the intermediately resistant mutants, and the magnitude of upregulation declined with the occurrence of mutations in the 23S rRNA gene. DNA microarray analysis revealed the differential expression of 265 genes, most of which occurred in the intermediate mutant, including the upregulation of genes encoding ribosomal proteins and the downregulation of genes involved in energy metabolism and motility. These results indicate (i) that mutations in L4 and L22 along with temporal overexpression of antibiotic efflux genes precede and may facilitate the development of high-level macrolide resistance and (ii) that the development of macrolide resistance affects the pathways important for physiology and metabolism inC. jejuni, providing an explanation for the reduced fitness of macrolide-resistantCampylobacter.

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Utility of Sequencing theerm(41) Gene in Isolates of Mycobacterium abscessus subsp. abscessus with Low and Intermediate Clarithromycin MICs

Journal of Clinical Microbiology ◽

10.1128/jcm.02950-14 ◽

2015 ◽

Vol 53 (4) ◽

pp. 1211-1215 ◽

Cited By ~ 84

Author(s):

Barbara A. Brown-Elliott ◽

Sruthi Vasireddy ◽

Ravikiran Vasireddy ◽

Elena Iakhiaeva ◽

Susan T. Howard ◽

...

Keyword(s):

Nucleotide Substitution ◽

Gene Mutations ◽

Mycobacterium Abscessus ◽

Macrolide Resistance ◽

23S Rrna ◽

Rrna Gene ◽

Content Type ◽

23S Rrna Gene ◽

Content Sequencing

Theerm(41) gene confers inducible macrolide resistance inMycobacterium abscessussubsp.abscessus, calling into question the usefulness of macrolides for treatingM. abscessussubsp.abscessusinfections. With an extended incubation (14 days), isolates with MICs of ≥8 μg/ml are considered macrolide resistant by current CLSI guidelines. Our goals were to determine the incidence of macrolide susceptibility in U.S. isolates, the validity of currently accepted MIC breakpoints, and theerm(41) sequences associated with susceptibility. Of 349 isolates (excluding those with 23S rRNA gene mutations), 85 (24%) had clarithromycin MICs of ≤8 μg/ml. Sequencing of theerm(41) genes from these isolates, as well as from isolates with MICs of ≥16 μg/ml, including ATCC 19977T, revealed 10 sequevars. The sequence in ATCC 19977Twas designated sequevar (type) 1; most macrolide-resistant isolates were of this type. Seven sequevars contained isolates with MICs of >16 μg/ml. The T28C substitution inerm(41), previously associated with macrolide susceptibility, was identified in 62 isolates (18%) comprising three sequevars, with MICs of ≤2 (80%), 4 (10%), and 8 (10%) μg/ml. No other nucleotide substitution was associated with macrolide susceptibility. We recommend that clarithromycin susceptibility breakpoints forM. abscessussubsp.abscessusbe changed from ≤2 to ≤4 μg/ml and that isolates with an MIC of 8 μg/ml have repeat MIC testing orermsequencing performed. Our studies suggest that macrolides are useful for treating approximately 20% of U.S. isolates ofM. abscessussubsp.abscessus. Sequencing of theermgene ofM. abscessussubsp.abscessuswill predict inducible macrolide susceptibility.

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Differences in the Frequency of 23S rRNA Gene Mutations in Mycoplasma pneumoniae between Children and Adults with Community-Acquired Pneumonia: Clinical Impact of Mutations Conferring Macrolide Resistance

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01421-12 ◽

2012 ◽

Vol 56 (12) ◽

pp. 6393-6396 ◽

Cited By ~ 24

Author(s):

Soo Jin Yoo ◽

Hyo-Bin Kim ◽

Sang-Ho Choi ◽

Sang-Oh Lee ◽

Sung-Han Kim ◽

...

Keyword(s):

Mycoplasma Pneumoniae ◽

Pediatric Patients ◽

Gene Mutations ◽

Community Acquired Pneumonia ◽

Clinical Impact ◽

Macrolide Resistance ◽

23S Rrna ◽

Rrna Gene ◽

Content Type ◽

23S Rrna Gene

ABSTRACTWe investigated the frequency and clinical significance of macrolide resistance in adult and pediatric patients with community-acquired pneumonia from aMycoplasma pneumoniaeinfection. The frequency of the A2063G mutation in the 23S rRNA gene was significantly higher in children than in adults (61.3% [19/31] and 13.3% [8/60], respectively;P< 0.001). Patients with macrolide-resistantM. pneumoniaeinfections showed a longer duration of fever (P= 0.021) and required a longer duration of antibiotic treatment (P= 0.007).

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Ribosomal Mutations as the Main Cause of Macrolide Resistance in Campylobacter jejuni and Campylobacter coli

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00314-11 ◽

2011 ◽

Vol 55 (12) ◽

pp. 5939-5941 ◽

Cited By ~ 29

Author(s):

Mirva Lehtopolku ◽

Pirkko Kotilainen ◽

Marjo Haanperä-Heikkinen ◽

Ulla-Maija Nakari ◽

Marja-Liisa Hänninen ◽

...

Keyword(s):

Macrolide Resistance ◽

23S Rrna ◽

Rrna Gene ◽

Campylobacter Coli ◽

Resistance Mutations ◽

Content Type ◽

Amino Acid Insertion ◽

23S Rrna Gene ◽

Ribosomal Protein L22 ◽

Resistant Strains

ABSTRACTThe aim of this study was to examine macrolide resistance mutations inCampylobacterspecies. In 76 strains studied, point mutation A to G at position 2059 of the 23S rRNA gene was detected in 30 of the 33 erythromycin-resistant strains. An amino acid insertion in the ribosomal protein L22 was found in one resistant strain without a 23S rRNA mutation. The A2059G mutation is the main cause of macrolide resistance inCampylobacterspecies.

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First Report of the 23S rRNA Gene A2058G Point Mutation Associated With Macrolide Resistance in Treponema pallidum From Syphilis Patients in Cuba

Sexually Transmitted Diseases ◽

10.1097/olq.0000000000000440 ◽

2016 ◽

Vol 43 (5) ◽

pp. 332-334 ◽

Cited By ~ 6

Author(s):

Angel A. Noda ◽

Nelvis Matos ◽

Orestes Blanco ◽

Islay Rodríguez ◽

Lola Virginia Stamm

Keyword(s):

Point Mutation ◽

Treponema Pallidum ◽

Macrolide Resistance ◽

23S Rrna ◽

Rrna Gene ◽

First Report ◽

23S Rrna Gene

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Mycoplasma genitalium acquisition and macrolide resistance after initiation of HIV pre-exposure prophylaxis in men who have sex with men

Sexually Transmitted Infections ◽

10.1136/sextrans-2019-054335 ◽

2020 ◽

Vol 96 (6) ◽

pp. 396-398 ◽

Cited By ~ 2

Author(s):

Jens Tomas Van Praet ◽

Sanne Steyaert ◽

Stefaan Vandecasteele ◽

Barbara Van Den Bergh ◽

Hilde Mahieu ◽

...

Keyword(s):

Mycoplasma Genitalium ◽

Macrolide Resistance ◽

23S Rrna ◽

Rrna Gene ◽

Younger Age ◽

23S Rrna Gene ◽

Sex With Men ◽

Taqman Array ◽

Exposure Prophylaxis

ObjectivesRecent evidence shows that patients using HIV pre-exposure prophylaxis (PrEP) have an increased rate of bacterial STIs, including syphilis, chlamydia and gonorrhoea. Our study aimed to describe the acquisition and the susceptibility for macrolides of Mycoplasma genitalium in men who have sex with men (MSM) on PrEP.MethodsWe studied all MSM who started PrEP in the AZ Sint-Jan Hospital Bruges from 1 June 2017 to 31 March 2019 with at least one follow-up visit. Patients were screened for M. genitalium and other STIs with pooled rectal swabs, pharyngeal swabs and first-voided urine, and blood samples at baseline and quarterly intervals after initiating PrEP. TaqMan Array Card technology was used to detect M. genitalium and determine macrolide-resistance mediating mutations in region V of the 23S rRNA gene (A2058G, A2059G, A2058C and others). Patients with an STI were treated based on a national guideline.Results131 MSM (median age 40 years, range 20–79) were included in the study. The median follow-up time was 12 months (IQR 6.1–17). Baseline prevalence of M. genitalium was 6.9% and incidence rate after PrEP initiation was 28.8 per 100 person-years (95% CI 21.7 to 37.2), without significant differences in proportions between the first four quarterly intervals. All but one acquisitions were asymptomatic. Younger age and positivity for M. genitalium at baseline were significantly associated with incident M. genitalium acquisition. The observed proportion of macrolide resistance increased not significantly from 44% at baseline to 57%–86% after PrEP initiation. None of the 27 macrolide-resistant M. genitalium acquisitions could be linked to azithromycin exposure in the three preceding months.ConclusionsAfter initiation of PrEP, we found a stable incidence of almost exclusively asymptomatic M. genitalium. However, a non-significant trend of an increased percentage of macrolide-resistant strains was observed.

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A 5′ Nuclease Genotyping Assay for Identification of Macrolide-Resistant Mycoplasma genitalium in Clinical Specimens

Journal of Clinical Microbiology ◽

10.1128/jcm.00012-16 ◽

2016 ◽

Vol 54 (6) ◽

pp. 1593-1597 ◽

Cited By ~ 15

Author(s):

Gitte Qvist Kristiansen ◽

Jan Gorm Lisby ◽

Kristian Schønning

Keyword(s):

Mycoplasma Genitalium ◽

Antimicrobial Treatment ◽

Macrolide Resistance ◽

23S Rrna ◽

Rrna Gene ◽

Predictive Values ◽

Content Type ◽

Clinical Specimens ◽

Genotyping Assay ◽

23S Rrna Gene

Rapid and sensitive detection of macrolide resistance inMycoplasma genitaliumis required for the guidance of adequate antimicrobial treatment. Previous studies have confirmed that single-base mutations at position 2058 or 2059 in domain V of the 23S rRNA gene ofM. genitaliumresult in high-level macrolide resistance. Sequencing of PCR products remains the gold standard for the identification of mutations conferring resistance to macrolides but is laborious and time-consuming. The aim of the present study was to develop a 5′ nuclease genotyping assay to detect single nucleotide polymorphisms in the 23S rRNA gene ofMycoplasma genitaliumthat are associated with macrolide resistance by combining PCR with hydrolysis probes and subsequent endpoint genotyping analysis. The 5′ nuclease genotyping assay was used as a referral test to be used onM. genitalium-positive samples and was validated on 259 positive samples, of which 253 (97.7%) were successfully sequenced. With the newly developed assay, 237/259 (91.5%) investigatedM. genitalium-positive samples were genotyped. The positive and the negative predictive values were 100% when evaluated on successfully genotyped samples. The newly developed assay discriminated macrolide-resistantM. genitaliumin clinical specimens possessing A2058G, A2058C, A2058T, and A2059G mutations with a sensitivity of 94.4% (95% confidence interval [CI], 90.7% to 98.2%) and a specificity of 92.7% (95% CI, 87.8% to 97.6%) when evaluated on successfully sequenced samples. The assay can correctly guide antimicrobial treatment ofM. genitaliuminfections.

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Molecular Basis of Macrolide Resistance inCampylobacterStrains Isolated from Poultry in South Korea

BioMed Research International ◽

10.1155/2018/4526576 ◽

2018 ◽

Vol 2018 ◽

pp. 1-9 ◽

Cited By ~ 3

Author(s):

Bai Wei ◽

Min Kang

Keyword(s):

Molecular Mechanisms ◽

Macrolide Resistance ◽

23S Rrna ◽

Rrna Gene ◽

Campylobacter Coli ◽

Erythromycin Resistance ◽

23S Rrna Gene ◽

Ribosomal Protein L22 ◽

Resistant Strains ◽

Azithromycin Resistance

We investigated the molecular mechanisms underlying macrolide resistance in 38 strains ofCampylobacterisolated from poultry. Twenty-seven strains were resistant to azithromycin and erythromycin, five showed intermediate azithromycin resistance and erythromycin susceptibility, and six showed azithromycin resistance and erythromycin susceptibility. FourCampylobacter jejuniand sixCampylobacter colistrains had azithromycin MICs which were 8–16 and 2–8-fold greater than those of erythromycin, respectively. The A2075G mutation in the 23S rRNA gene was detected in 11 resistant strains with MICs ranging from 64 to ≥ 512μg/mL. Mutations including V137A, V137S, and a six-amino acid insertion (114-VAKKAP-115) in ribosomal protein L22 were detected in theC. jejunistrains. Erythromycin ribosome methylase B-erm(B) was not detected in any strain. All strains except three showed increased susceptibility to erythromycin with twofold to 256-fold MIC change in the presence of phenylalanine arginine ß-naphthylamide (PAßN); the effects of PAßN on azithromycin MICs were limited in comparison to those on erythromycin MICs, and 13 strains showed no azithromycin MIC change in the presence of PAßN. Differences between azithromycin and erythromycin resistance and macrolide resistance phenotypes and genotypes were observed even in highly resistant strains. Further studies are required to better understand macrolide resistance inCampylobacter.

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