Erm Gene Research Articles

Detection of genes conferring resistance on multi antibiotic resistance bacteria isolated from ready to eat foods sold in the Nigerian defence academy, Kaduna, Nigeria

Food has always been the means by which disease-causing organisms are transmitted to man. This study sought to isolate multi-antibiotic resistant bacteria from ready to eat food (RTE) sold within Nigerian Defence Academy. Eight (8) food samples were collected from five (5) vendors and spread plate technique was adopted for bacteria isolation from the food samples. Twenty-five (25) bacteria isolates were obtained from the food samples and subjected to antibiotic sensitivity assay using Kirby Bauer disc diffusion method. Eight (8) of the isolates exhibited various degrees of resistance to the tested antibiotics. All the isolated bacteria in this study were susceptible to the effect of Quinolones antibiotics and were resistant to cefuroxime, cefotaxime and Ampiclox. Six (6) of the isolates that showed multi-antibiotic resistance were characterized by 16SrRNA gene sequencing as Bacillus cereus strains (0083 and HYM76), Staphylococcus aureus strain MRSA252, Lactobacillus murinus strain AMB_10 and Listeria Spp. Polymerase chain reaction (PCR) and agarose gel electrophoresis detection of four (4) antibiotics resistance genes – Aminoglycoside adenylyltransferase (AADA1), Acridine (ACRA), Chloramphenicol Resistance (CMLA) and Erythromycin (ERM(C)) – produced the following results: AADA1 genes were amplified in Staphylococcus aureus and Bacillus cereus; ACRA gene was detected in all the selected isolates; CMLA gene was detected in Staphylococcus aureus, Lactobacillus murinus and Bacillus cereus while ERM(C) gene was present in Staphylococcus aureus.

Molecular characterization of the multi-drug resistant Myroides odoratimimus isolates: a whole genome sequence-based study to confirm carbapenem resistance.

The bacteria belonging to the Myroides genus are opportunistic pathogens causing community or hospital-acquired infections that result in treatment failure due to antibiotic resistance. This study aimed to investigate molecular mechanisms of antibiotic resistance, clonal relatedness, and the biofilm forming capacity of the 51 multi-drug resistant Myroides odoratimimus. All isolates were screened for blaKPC, blaOXA, blaVIM, blaIMP, blaMUS, blaTUS, blaNDM, and blaB genes by using PCR amplification. Whole genome sequencing (WGS) was applied on three randomly selected isolates for further investigation of antibiotic resistance mechanisms. Clonal relatedness was analyzed by Pulsed-field gel electrophoresis (PFGE) and the microtiter plate method was used to demonstrate biofilm formation. All isolates were positive for biofilm formation. PCR analysis resulted in a positive for only the blaMUS-1 gene. WGS identified blaMUS-1, erm(F), ere(D), tet(X), and sul2 genes in all strains tested. Moreover, the genomic analyses of three strains revealed that genomes contained a large number of virulence factors (VFs). PFGE yielded a clustering rate of 96%. High clonal relatedness, biofilm formation, and multi-drug resistance properties may lead to the predominance of these opportunistic pathogens in hospital environments and make them cause nosocomial infections.

Prevalence and Molecular Characterization of Macrolide Resistance in Clinical Isolates of Beta Hemolytic Streptococci from a Tertiary Care Teaching Hospital in Kerala, India

Background: Beta-hemolytic streptococci (BHS) are responsible for both invasive and noninvasive infections, and the preferred treatment for these infections is penicillin due to the distinctive characteristics of these bacteria. However, in patients who cannot tolerate β-lactam antibiotics, macrolides and clindamycin are important alternative options for treating BHS infections. This study aimed to analyse the pattern of macrolide resistance among clinical strains of beta-hemolytic streptococci (BHS).
 Methods: Beta hemolytic streptococci isolated from clinical specimens during December 2018 to May 2020 were included in this study. Identification of the isolates were done by conventional and Vitek 2 method. All isolates were subjected to serogrouping. Antibiotic susceptibility testing done by disc diffusion method. Genes encoding macrolide resistance were detected by conventional multiplex polymerase chain reaction.
 Results: A total of 129 beta hemolytic streptococcal isolates were obtained which included 27 S. pyogenes (20.9%), 77 S.agalactiae (59.7%), 23 S.dysgalactiae spp equisimilis (17.8%) and one isolate of S.anginosus and S. porcinus each (0.8%). Erythromycin, clindamycin, quinuprisitn and tetracycline resistance were found to be 20.2%, 12.1%, 16.3% and 51.2% respectively. Among the 26 erythromycin resistant isolates, 12(46.2%) were inducible clindamycin resistant phenotype. Out of 26 erythromycin resistant isolates, 7(26.9%) isolates were harbouring erm(A) gene, 10(38.5%) erm(B) and 9(34.6%) mef(A) gene.
 Conclusion: Our study highlights the importance of routine antibiotic susceptibility testing for betahaemolytic streptococci, as well as the detection of inducible resistance to prevent therapeutic failure.

Multiplex Real-Time PCR for the Detection of Tetracycline, Ciprofloxacin, and Erythromycin Resistance Determinants from Human and Foodborne Campylobacter jejuni and Campylobacter coli.

Campylobacter jejuni and Campylobacter coli are the predominant thermophilic species responsible for foodborne gastroenteritis worldwide. Elevated resistance to certain antibiotics was observed due to antimicrobial therapy in farm animals and humans, while reduced antimicrobial usage partially reduced antibiotic resistance. Monitoring the antimicrobial resistance demonstrated a substantial fraction of multi-resistant isolates, indicating the necessity of reliable tools for their detection. In this study, resistance determinants in 129 German and 21 Vietnamese isolates were selected to establish a novel multiplex real-time PCR (qPCR), facilitating the simultaneous detection of four resistance determinants. These comprised tet(O) gene variants associated with tetracycline resistance, point mutations GyrA_T86I and GyrA_T86V associated with ciprofloxacin resistance, and the erm(B) gene together with the point mutation A2075G in the 23S rRNA gene, associated with erythromycin resistance. Moreover, the performance of the qPCR assay was evaluated by comparing the results of qPCR to phenotypic antimicrobial resistance profiles, obtained with standardized EUCAMP3 microdilution panel, which showed 100% similarity (inclusivity and exclusivity). Variation in measurement methods, including qPCR machines and master mixes showed robustness, essential for laboratories. The assay can be used for the rapid detection of resistance determinants, and is beneficial for monitoring the spread of antibiotic resistance in C. jejuni and C. coli.

Microbial community of municipal drinking water in Hangzhou using metagenomic sequencing

While traditional culture-dependent methods can effectively detect certain microorganisms, the comprehensive composition of the municipal drinking water (DW) microbiome, including bacteria, archaea, and viruses, remains unknown. Metagenomic sequencing has opened the door to accurately determine and analyze the entire microbial community of DW, providing a comprehensive understanding of DW species diversity, especially in the context of public health concerns during the COVID-19 era. In this study, we found that most of the culturable bacteria and some fecal indicator bacteria, such as Escherichia coli and Pseudomonas aeruginosa, were non-culturable using culture-dependent methods in all samples. However, metagenomic analysis showed that the predominant bacterial species in the DW samples belonged to the phyla Proteobacteria and Planctomycetes. Notably, the genus Methylobacterium was the most abundant in all water samples, followed by Sphingomonas, Gemmata, and Azospirilum. While low levels of virulence-associated factors, such as the Esx-5 type VII secretion system (T7SS) and DevR/S, were detected, only the erythromycin resistance gene erm(X), an rRNA methyltransferase, was identified at low abundance in one sample. Hosts corresponding to virulence and resistance genes were identified in some samples, including Mycobacterium spp. Archaeal DNA (Euryarchaeota, Crenarchaeota) was found in trace amounts in some DW samples. Viruses such as rotavirus, coxsackievirus, human enterovirus, and SARS-CoV-2 were negative in all DW samples using colloidal gold and real-time reverse transcription polymerase chain reaction (RT‒PCR) methods. However, DNA encoding a new order of reverse-transcribing viruses (Ortervirales) and Herpesvirales was found in some DW samples. The metabolic pathways of the entire microbial community involve cell‒cell communication and signal secretion, contributing to cooperation between different microbial populations in the water. This study provides insight into the microbial community and metabolic process of DW in Hangzhou, China, utilizing both culture-dependent methods and metagenomic sequencing combined with bioinformatics tools during the COVID-19 pandemic era.

2459. Prevalence of Nasopharyngeal Carriage of Macrolide and Lincosamide Resistance Genes Among a Household-based cohort of Children and Adults in a Peri-urban Community in Peru

Abstract Background Antibiotic resistant infections commonly occur in healthcare settings, but the prevalence of antibiotic resistant genes (ARGs) in healthy individuals in the community is unknown. We provide an initial assessment of nasopharyngeal carriage of ARGs conferring resistance to macrolides and/or lincosamides, including erm, mph, ABC-F, and lnu genes. Methods Nasopharyngeal swabs were systematically obtained at enrollment and weekly thereafter from children and adults enrolled in a household-based prospective cohort study in Lima, Peru. Samples were sequenced using the Illumina Respiratory Pathogen/ID AMR Panel to detect common respiratory bacteria and ARGs. We defined ‘any erm gene’ (erm) as the detection of any specific erm gene class, ‘any mph gene’ as detection of any specific mph gene class, ‘any ABC-F gene’ as any specific ABC-F gene class, and ‘any lnu gene’ as any specific lnu gene detection. We compared the prevalence of erm, mph, ABC-F and lnu gene colonization at enrollment among age groups (ages 0-4, 5-17, 18-44, and 45+ years) using the Fisher's exact test. We then compared the frequency of co-detection of two or more macrolide or lincosamide ARGs between both pediatric age groups (ages 0-17 years) and both adult age groups (18+ years). Results 114 individuals were included in this analysis (Table 1). At least one macrolide or lincosamide ARG was detected in 61/114 (53.5%) individuals. The prevalence of erm, mph, ABC-F, or lnu gene detection was similar among age groups (Figure 1). Two or more genes were co-detected in 45/114 (39.4%) individuals. Co-detections with two or more erm, mph, ABC-F, or lnu genes were more frequent in children 0-17 years compared to adults 18+ years (p=0.013) (Figure 2). Conclusion Conclusions: These preliminary results indicate that macrolide or lincosamide ARGs were commonly detected in healthy community-dwelling children and adults in Lima, Peru. The prevalence of erm, mph, ABC-F, and lnu genes was similar across age groups, and co-detection of ARGs was more common in children. Future studies will assess changes in ARG carriage over time, transmission among household members, and its clinical relevance. Disclosures Claudio Lanata, MD, MPH, CureVAc AG Germany: Grant/Research Support|HilleVac Inc.: Advisor/Consultant|HilleVac Inc.: Grant/Research Support Carlos G. Grijalva, MD, MPH, AHRQ: Grant/Research Support|CDC: Grant/Research Support|FDA: Grant/Research Support|Merck: Advisor/Consultant|NIH: Grant/Research Support|Syneos Health: Grant/Research Support

First Report of Plasmid-Mediated Macrolide-Clindamycin-Tetracycline Resistance in a High Virulent Isolate of Cutibacterium acnes ST115.

Cutibacterium acnes, a prevalent skin commensal, has emerged as a significant global challenge due to its widespread antibiotic resistance. To investigate the antibiotic resistance mechanisms and clinical characterization of C. acnes in Korea, we collected 22 clinical isolates from diverse patient specimens obtained from the National Culture Collection for Pathogens across Korea. Among the isolates, KB112 isolate was subjected to whole genome sequencing due to high resistance against clindamycin, erythromycin, tetracycline, doxycycline, and minocycline. The whole genome analysis of KB112 isolate revealed a circular chromosome of 2,534,481 base pair with an average G + C content of 60.2% with sequence type (ST) 115, harboring the potential virulent CAMP factor pore-forming toxin 2 (CAMP2), the multidrug resistance ABC transporter ATP-binding protein YknY, and the multidrug efflux protein YfmO. The genomic sequence also showed the existence of a plasmid (30,947 bp) containing the erm(50) and tet(W) gene, which confer resistance to macrolide-clindamycin and tetracycline, respectively. This study reports plasmid-mediated multi-drug resistance of C. acnes for the first time in Korea.

Aortic endograft infection by Mycobacterium abscessus subsp. massiliense with acquired clarithromycin resistance: a case report

BackgroundMycobacterium abscessus subsp. massiliense (MMA) comprises a group of non-tuberculous, rapidly growing mycobacteria. Although MMA can cause pulmonary diseases, surgical site infections, and disseminated diseases, aortic endograft infection has not been reported. Here, we describe the first case of aortic endograft infection caused by MMA.Case presentationTwo months after stent-graft insertion for an abdominal aortic aneurysm, an 85-year-old man was admitted with fever and abdominal pain and was diagnosed with aortic endograft infection. Despite 14 days of meropenem and vancomycin intravenous administration, periaortic fluid pooling increased as compared to that before antibiotic administration. The abscess was drained, and fluorescent acid-fast staining of the abscess fluid revealed bacilli. We conducted genetic tests on the genes hsp65, rpoB, and sodA, performed Whole Genome Sequencing (WGS), and identified the organism as MMA. Intravenous imipenem–cilastatin (IPM/CS), amikacin (AMK), and oral clarithromycin (CAM) were administered. After 2 months, oral CAM and sitafloxacin were administered because the abscess had decreased in size. However, after 6 weeks, the abscess increased in size again. Antimicrobial susceptibility testing of the drainage fluid from the abscess resulted in the isolation of an MMA strain that had acquired resistance to CAM. Intravenous IPM/CS, AMK, and oral linezolid were added to the treatment regimen along with oral CAM and STFX. However, he was not fully cured and died 6 months later. Neither the full-length erythromycin ribosome methyltransferase (erm)(41) gene nor the rrl or rpIV gene mutations were found by Sanger sequencing in the pre- and post-treatment strains. Whole-genome sequence analysis of the post-treatment strain revealed mutations in genes with no previous reports of association with macrolide resistance.ConclusionsAortic endograft infection caused by MMA strain is extremely rare; nonetheless, MMA should be suspected as the causative microorganism when broad-spectrum antimicrobials are ineffective.

Characterization of a non-functional erm(T) gene in a clinical Enterococcus faecium strain.

Characterization of a non-functional erm(T) gene in a clinical Enterococcus faecium strain.

Macrolide-Lincosamide Resistance and Virulence Genes in Staphylococcus aureus Isolated from Clinical Specimens in Ardabil, Iran.

Staphylococcus aureus causes various hospital- and community-acquired infections. This study aimed to investigate the phenotypic and genotypic characteristics of erythromycin and inducible clindamycin resistance, virulence gene profiles, and spa types of S. aureus isolates collected from patients in Ardabil Province, Iran. A total of 118 clinical S. aureus isolates, including 50 (42.4%) methicillin-resistant S. aureus (MRSA) and 68 (57.6%) methicillin-susceptible S. aureus (MSSA) strains, were investigated. Resistance patterns were determined by the disk diffusion method and minimum inhibitory concentration (MIC) test. Inducible macrolide-lincosamide-streptogramin B (iMLSB) resistance was detected using D-test method. The polymerase chain reaction (PCR) was used to identify the virulence and resistance-encoding genes. Additionally, the spa types of the isolates were determined using the PCR, followed by sequencing. In total, 49.1% (58/118) and 44% (52/118) of the isolates were resistant to erythromycin and clindamycin, respectively. Overall, 13.5% (16/118) of the isolates showed the iMLSB resistance phenotype. The ermC gene (72.4% [42]) was the most frequent erythromycin resistance-encoding gene, followed by ermA (60.3% [35]), ermB (60.3% [35]), ermTR (51.7% [30]), and msrA (15.5% [9]) genes among erythromycin-resistant isolates. The virulence genes hla, hld, sea, LukS PV, tst, seb, sed, eta, sec, and etb were detected in 93.2%, 74.5%, 70.3%, 32.2%, 29.6%, 17%, 8.5%, 8.5%, 5.9%, and 4.2% of the isolates, respectively. Ten different spa types were identified for 58 erythromycin-resistant S. aureus strains, of which t030 and t078 types were the most common types. A high frequency of macrolide- and lincosamide-resistant S. aureus isolates with different genetic backgrounds of resistance and virulence may be found in patients in Ardabil Province, Iran.

An increase in erythromycin resistance in methicillin-susceptible Staphylococcus aureus from blood correlates with the use of macrolide/lincosamide/streptogramin antibiotics. EARS-Net Spain (2004-2020).

To describe and analyse erythromycin resistance trends in blood isolates of Staphylococcus aureus (EARS-Net Spain, 2004-2020) and the association of these trends with the consumption of macrolide, lincosamide, and streptogramin B (MLSB) antibiotics. To assess molecular changes that could be involved in erythromycin resistance trends by whole genome analysis of representative isolates. We collected antibiotic susceptibility data for all first-blood S. aureus isolates in patients from 47 Spanish hospitals according to EARS-Net criteria. MLSB antibiotic consumption was obtained from the Spanish Agency for Medicines and Medical Devices (2008-2020). We sequenced 137 representative isolates for core genome multilocus sequence typing, resistome and virulome analysis. For the 36,612 invasive S. aureus isolates, methicillin resistance decreased from 26.4% in 2004 to 22.4% in 2020. Erythromycin resistance in methicillin-susceptible S. aureus (MSSA) increased from 13.6% in 2004 to 28.9% in 2020 (p < 0.001); however, it decreased from 68.7 to 61.8% (p < 0.0001) in methicillin-resistant S. aureus (MRSA). Total consumption of MLSB antibiotics increased from 2.72 defined daily doses per 1,000 inhabitants per day (DID) in 2014 to 3.24 DID in 2016. By WGS, the macrolide resistance genes detected were erm (59.8%), msrA (46%), and mphC (45.2%). The erm genes were more prevalent in MSSA (44/57, 77.2%) than in MRSA (38/80, 47.5%). Most of the erm genes identified in MSSA after 2013 differed from the predominant ermC gene (17/22, 77.3%), largely because ermT was significantly associated with MSSA after 2013 (11/29, 37.9%). All 13 ermT isolates in this study, except one, belonged to ST398 and came from 10 hospitals and six Spanish provinces. The significant increase in erythromycin resistance in blood MSSA correlated with the consumption of the MLSB antibiotics in Spain. These preliminary data seem support the hypothesis that the human ST398 MSSA clade with ermT-mediated resistance to erythromycin may be involved in this trend.

Plasmid-mediated drug resistance in mycobacteria: the tip of the iceberg?

Macrolides, such as clarithromycin, are crucial in the treatment of nontuberculous mycobacteria (NTM). NTM are notoriously innately drug resistant, which has made the dependence on macrolides for their treatment even more important. Not surprisingly, resistance to macrolides has been documented in some NTM, including Mycobacterium avium and Mycobacterium abscessus, which are the two NTM species most often identified in clinical isolates. Resistance is mediated by point mutations in the 23S ribosomal RNA or by methylation of the rRNA by a methylase (encoded by an erm gene). Chromosomally encoded erm genes have been identified in many of the macrolide-resistant isolates, but not in Mycobacterium chelonae. Now, Brown-Elliott et al. (J Clin Microbiol 61:e00428-23, 2023, https://doi.org/10.1128/JCM.00428-23) describe the identification of a new erm variant, erm(55), which was found either on the chromosome or on a plasmid in highly macrolide-resistant clinical isolates of M. chelonae. The chromosomal erm(55) gene appears to be associated with mobile elements; one gene is within a putative transposon and the second is in a large (37 kb) insertion/deletion. The plasmid carrying erm(55) also encodes type IV and type VII secretion systems, which are often linked on large mycobacterial plasmids and are hypothesized to mediate plasmid transfer. While the conjugative transfer of the erm(55)-containing plasmid between NTM has yet to be demonstrated, the inferences are clear, as evidenced by the dissemination of plasmid-mediated drug resistance in other medically important bacteria. Here, we discuss the findings of Brown-Elliott et al., and the potential ramifications on treatment of NTM infections.

Distribution of MecA and Erm Genes among Methicillin-resistant Staphylococcus Aureus with Inducible Resistance to Clindamycin.

The emergence of Methicillin-resistant Staphylococcus aureus and its ability to confer cross-resistance to macrolide-lincosamide-streptogramin B has complicated the treatment against it. Gene-based studies among phenotypic methicillin-resistant isolates with inducible resistance to clindamycin are less available in Nepal. This work was undertaken to detect the mecA and erm genes among such phenotypes isolated from clinical samples. S. aureus isolated from different clinical samples was identified by standard microbiological procedures (Gram-staining, colony morphology, and different biochemical tests). Methicillin-resistant and inducible resistant to clindamycin phenotypes were detected by using cefoxitin disc (30 µg) and a double disk diffusion test according to the Clinical and Laboratory Standards Institute guidelines and mecA and erm genes were detected by polymerase chain reaction. Among 120 S. aureus isolates, 51.67% (n=62) were MRSA, and the prevalence of inducibly-resistant, constitutively-resistant and Macrolide-Streptogramin phenotypes were 15.83% (n=19), 28.33% (n=34) and 15.83% (n=19) respectively. While 35.84% (n=43) of isolates showed sensitivity to both antibiotics, erythromycin and clindamycin. Out of 14 inducibly-resistant phenotypes, 57.14% (n=8) were found carrying ermC and 28.57% (n=4) phenotypes contained both ermA and ermC. All phenotypes were positive for the mecA gene. Macrolides-Lincosamide-Streptogramin B resistance was predominant among methicillin-resistant S. aureus. While all isolates with inducible clindamycin resistance harbored mecA gene, most of them also harbored ermC gene. The higher prevalence of inducible-resistant to clindamycin indicated the need for rational use of antimicrobial agents.

Distribution of Macrolide, Lincosamide, and Streptogramin B and Detection of erm Genes in Staphylococcus aureus from Wounds in Uyo, Nigeria

The increasing resistance to macrolide, lincosamide, and streptogramin B (MLSB) among methicillin-resistant Staphylococcus aureus (MRSA) is a challenge. The study determined the constitutive and inducible MLSB resistance and erm genes in S. aureus from wounds using the erythromycin-clindamycin D-zone test and a multiplex polymerase chain reaction assay, respectively. Of the 260 patients recruited in the study, S. aureus was isolated from the wounds of 102 patients, giving a prevalence rate of 39.2%. Of the 102 S. aureus isolates, 32.4% were methicillin-sensitive S. aureus (MSSA), and 67.6% were MRSA. The prevalence of S. aureus in wounds was higher in females (39.8%) than in males (38.4%). The highest prevalence of S. aureus was found in divorce patients (59.1%) and those residing in urban areas (39.7%). There was no statistical difference between the occurrence of MRSA and MSSA in wounds based on the sex (p = 0.97), age (p = 0.08), and marital status (p = 0.41) of the patients. All (100%) MRSA were resistant to Cefoxitin, and chloramphenicol, while ˂ 50% of MRSA were resistant to Tetracycline, Ciprofloxacin, and Gentamycin. Of the 69 MRSA isolates from the wounds, 21.7% and 26.1% were iMLSB and cMLSB phenotypes, respectively. Nine (9) MSSA were iMLSB phenotypes, and eight MSSA were cMLSB phenotypes. Among the 12 representative isolates, three (3) MRSA and one (1) MSSA isolate possessed the ermC gene. This study has revealed that screening tests for iMLSB-resistant S. aureus strains are critical for the therapeutic management of wound infections caused by S. aureus.

Prevalence of erythromycin-resistant emm92-type invasive group A streptococcal infections among injection drug users in West Virginia, United States, 2021-23.

Increasing incidence of invasive group A Streptococcus (iGAS) disease has been reported in Europe and the USA over the past several years. Coupled with this are observations of higher rates of resistance to erythromycin and clindamycin. To characterize iGAS and pharyngitis isolates from West Virginia (WV), a US state outside of the national Active Bacteria Core surveillance purview, where risk factors associated with iGAS infections are prevalent. Seventy-seven invasive group A Streptococcus isolates were collected from 67 unique patients at the J.W. Ruby Memorial Hospital Clinical Microbiology Laboratory in WV from 2021 to 2023. Invasive isolates and 20 unique pharyngitis isolates were tested for clindamycin and erythromycin susceptibility in the clinical laboratory. Patient demographic and clinical information was retrieved from patient electronic health records. Isolates were further characterized based on emm subtype and detection of MLSB resistance determinants. Twenty-six (39%) isolates were of a single emm92 type. All emm92 isolates were uniformly erythromycin/clindamycin resistant with inducible or constitutive MLSB resistance imparted by the plasmid-borne erm(T) gene. The majority of emm92 infections were associated with adult patients who reported IV drug use, whereas no pharyngitis infections were caused by an emm92 strain. Overall, 51 (76%) of the 67 iGAS isolates were determined to carry MLSB resistance. Isolates of emm92 type (clonal subtype emm92.0) were associated with iGAS infections in adult IV drug users, but not with paediatric pharyngitis, and were uniformly resistant to erythromycin and clindamycin.

Unusually high clarithromycin resistance in Mycobacterium abscessus subsp. abscessus isolated from human gastric epithelium.

Mycobacterium abscessus subsp. abscessus is a rapidly growing facultative intracellular pathogen that usually infects human lung and skin epithelium. Recently, we and another group have shown that it also has the potential to colonize human gastric epithelium, but its significance with respect to gastric diseases remains unclear. Although Helicobacter pylori still remains the only definite gastric pathogen, recent studies have shown that M. abscessus subsp. abscessus also has the potential to colonize human gastric epithelium. M. abscessus subsp. abscessus is known to exhibit multidrug resistance and clarithromycin has been used as the drug of choice. We aimed to determine the clarithromycin resistance profile of 117 (74 rough and 43 smooth) gastric M. abscessus subsp. abscessus strains and to detect the point mutations in rrl and erm (41) genes conferring the resistance. Our data showed 79.48% (19 smooth and 74 rough) of M. abscessus subsp. abscessus strains were resistant to clarithromycin (MIC90 ≤ 512 μg/mL), while 20.51% (24 smooth) were susceptible (MIC90 ≤ 8 μg/mL). Nucleotide sequence analysis of the rrl gene with reference strains of M. abscessus subsp. abscessus did not show any mutation that is relevant to the clarithromycin resistance. However, analysis of erm (41) gene showed that M. abscessus subsp. abscessus strains, which were susceptible to clarithromycin had C, C, G, and C at their nucleotide positions 28, 159, 238, and 330, respectively, while the resistant strains showed T, T, A, and A at the same positions. Based on antibiogram and sequence analysis data we recommend further studies involving genomic analysis to identify the other genes involved in high clarithromycin resistance in gastric M. abscessus subsp. abscessus along with the mechanisms involved.

Novel macrolide-lincosamide-streptogramin B resistance gene erm(56) in Trueperella pyogenes.

Whole-genome sequence analysis of a macrolide, lincosamide, streptogramin B (MLSB)-resistant Trueperella pyogenes from a dog revealed a new 23S ribosomal RNA methylase gene erm(56). Expression of the cloned erm(56) confers resistance to MLSB in T. pyogenes and Escherichia coli. The erm(56) gene was flanked by two IS6100 integrated on the chromosome next to a sul1-containing class 1 integron. GenBank query revealed additional erm(56)-containing elements in another T. pyogenes and in Rothia nasimurium from livestock. IMPORTANCE A novel 23S ribosomal RNA methylase gene erm(56) flanked by insertion sequence IS6100 was identified in a Trueperella pyogenes isolated from the abscess of a dog and was also present in another T. pyogenes and in Rothia nasimurium from livestock. It was shown to confer resistance to macrolide, lincosamide, streptogramin B antibiotics in T. pyogenes and E. coli, indicating functionality in both Gram-positive and Gram-negative bacteria. The detection of erm(56) on different elements in unrelated bacteria from different animal sources and geographical origins suggests that it has been independently acquired and likely selected by the use of antibiotics in animals.

Baseline azithromycin resistance in the gut microbiota of preterm born infants

BackgroundMacrolides, including azithromycin, are increasingly used in preterm-born infants to treat Ureaplasma infections. The baseline carriage of macrolide resistance genes in the preterm stool microbiota is unknown.ObjectivesIdentify carriage of azithromycin resistant bacteria and the incidence of macrolide resistant genes.MethodsAzithromycin resistant bacteria were isolated from serial stool samples obtained from preterm infants (≤32 weeks’ gestation) by culturing aerobically/anaerobically, in the presence/absence of azithromycin. Using quantitative PCR, we targeted 6 common macrolide resistance genes (erm(A), erm(B), erm(C), erm(F), mef(A/E), msr(A)) in DNA extracted from selected bacteria resistant to azithromycin.ResultsFrom 89 stool samples from 37 preterm-born infants, 93.3% showed bacterial growth in aerobic or anaerobic conditions. From the 280 azithromycin resistant isolates that were identified, Staphylococcus (75%) and Enterococcus (15%) species dominated. Macrolide resistance genes were identified in 91% of resistant isolates: commonest were erm(C) (46% of isolates) and msr(A) (40%). Multiple macrolide resistance genes were identified in 18% of isolates.ConclusionMacrolide resistance is common in the gut microbiota of preterm-born infants early in life, most likely acquired from exposure to the maternal microbiota. It will be important to assess modulation of macrolide resistance, if macrolide treatment becomes routine in the management of preterm infants.Impact StatementAzithromycin resistance is present in the stool microbiota in the first month of life in preterm infants91% of azithromycin resistant bacteria carried at least one of 6 common macrolide resistant genesIncreasing use of macrolides in the preterm population makes this an important area of study

Using of Modern Genetic Methods to Detect Air-Borne Bacterial Genes Through Desert Storms in Iraq

Abstract Antibiotic resistance of pathogens is a serious issue that affects health. The airborne particle molecules such as (PM2.5, PM10) that spread by dust storms can increase the antibiotic resistance in bacterial community. In this study air samples of the dust storms were collected and the air-borne bacteria were cultured on specific growth media by portable air sampler device (SAS). Sixty plates were used through the atmospheric storms dust which included three periods of 24/5/2022,13/6/2022 and 3/7/2022. Antibiotic resistance of bacterial isolates was detected by VITEK2. Bacterial whole DNA was extracted and the genes against the most resisted antibiotics were detected by specific pre-designed primers and by using gel electrophoresis method. The results showed homogenous high concentrations of both PM2.5 and PM10 were included in storm samples just like that in Saudi Arabia. The greatest numbers of bacterial growth in collecting plates through the storm included 45 of 60 (75%) E. coli, 36 of 60 (60%) of Staphylococcus epidermis and 15 of 60 (25%) of Sphingomonas paucimobilis isolates of total bacterial growth as diagnosed by morphological characteristics and VITEK2 system. The antibiotic sensitivity test (AST) which was done by VITIK2 showed multidrug resistant for Staphylococcus epidermis, E. coli and Sphingomonas paucimobilis strains, mainly against erythromycin and tertacycline. The results of genetic studies for the antibiotic resistance illustrated the presence of gene fragments of both ermA gene and tetK gene within the bacterial nucleic acid, which are considered as potent antibiotic resistance genes (ARGs genes). In conclusion, the resistance of air-borne bacteria towards used antibiotics were elevated strongly in relation with the expression of ARGs associated with the huge amounts of PM particles in dust storms that occurred in Iraq, which is very harmful for human and other living forms on earth.

Structural insights into the mechanism of overcoming Erm-mediated resistance by macrolides acting together with hygromycin-A

The ever-growing rise of antibiotic resistance among bacterial pathogens is one of the top healthcare threats today. Although combination antibiotic therapies represent a potential approach to more efficiently combat infections caused by susceptible and drug-resistant bacteria, only a few known drug pairs exhibit synergy/cooperativity in killing bacteria. Here, we discover that well-known ribosomal antibiotics, hygromycin A (HygA) and macrolides, which target peptidyl transferase center and peptide exit tunnel, respectively, can act cooperatively against susceptible and drug-resistant bacteria. Remarkably, HygA slows down macrolide dissociation from the ribosome by 60-fold and enhances the otherwise weak antimicrobial activity of the newest-generation macrolide drugs known as ketolides against macrolide-resistant bacteria. By determining a set of high-resolution X-ray crystal structures of drug-sensitive wild-type and macrolide-resistant Erm-methylated 70S ribosomes in complex with three HygA-macrolide pairs, we provide a structural rationale for the binding cooperativity of these drugs and also uncover the molecular mechanism of overcoming Erm-type resistance by macrolides acting together with hygromycin A. Altogether our structural, biochemical, and microbiological findings lay the foundation for the subsequent development of synergistic antibiotic tandems with improved bactericidal properties against drug-resistant pathogens, including those expressing erm genes.