1887

Abstract

Limited information is available regarding the genomic characteristics of causing ear infections. Our aim is to characterize the genotypic features of an emerging ST316 sublineage causing aural infections in Shanghai. A total of 199 ear swab isolates were subjected to whole genome sequencing (WGS). Complete genomes for two isolates were resolved. We showed this recently emerged sublineage exhibited high-level resistance to fluoroquinolones (FQs) primarily by accumulation of known mutations in quinolone resistance determining regions (QRDRs). Loss-of-function mutations in and were frequently detected. Mutations in (P166S) and (S492F) were resident in this sublinage about 2 years after its emergence. Recombination events might be a key driver of genomic diversity in this sublineage. Convergent evolution events on Multidrug-resistant (MDR) determinants were also observed. We generated predictive machine models and identified biomarkers of resistance to gentamicin, fosfomycin, and cefoperazone-sulbactam in this sublineage. This sublineage tended to be less virulent by loss of a series virulence genes represented by , and iron uptake- and antimicrobial activity-related genes. Specific mutations were detected in and genes that related to surface structures. Moreover, this sublineage differed from non-ST316 isolates in several ways, including virulence genes related to cell surface structure. Our analysis suggested acquisition of a roughly 390 kbp MDR plasmid carrying might play an important role in the success of this sublinage. Clonal expansion of this sublineage exhibiting enhanced adaptation to cause ear infections is concerning, which requires urgent control measures to be implemented.

Funding
This study was supported by the:
  • Shanghai Municipal Science and Technology Commission (Award 19JC1413002)
    • Principle Award Recipient: BaixingDing
  • National Natural Science Foundation of China (Award 81902103)
    • Principle Award Recipient: BaixingDing
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License.
Loading

Article metrics loading...

/content/journal/mgen/10.1099/mgen.0.000989
2023-04-20
2024-05-04
Loading full text...

Full text loading...

/deliver/fulltext/mgen/9/4/mgen000989.html?itemId=/content/journal/mgen/10.1099/mgen.0.000989&mimeType=html&fmt=ahah

References

  1. Fischer S, Dethlefsen S, Klockgether J, Tümmler B. Phenotypic and genomic comparison of the two most common exoU-positive Pseudomonas aeruginosa clones, PA14 and ST235. mSystems 2020; 5:e01007-20 [View Article] [PubMed]
    [Google Scholar]
  2. Perveen S, Naqvi SB, Fatima A. Antimicrobial susceptibility pattern of clinical isolates from cases of ear infection using amoxicillin and cefepime. Springerplus 2013; 2:288 [View Article] [PubMed]
    [Google Scholar]
  3. Rubin Grandis J, Branstetter BF 4th, Yu VL. The changing face of malignant (necrotising) external otitis: clinical, radiological, and anatomic correlations. Lancet Infect Dis 2004; 4:34–39 [View Article] [PubMed]
    [Google Scholar]
  4. Mittal R, Lisi CV, Kumari H, Grati M, Blackwelder P et al. Otopathogenic Pseudomonas aeruginosa enters and survives inside macrophages. Front Microbiol 2016; 7:1828 [View Article] [PubMed]
    [Google Scholar]
  5. Livermore DM. Multiple mechanisms of antimicrobial resistance in Pseudomonas aeruginosa: our worst nightmare?. Clin Infect Dis 2002; 34:634–640 [View Article] [PubMed]
    [Google Scholar]
  6. Hwang W, Yong JH, Min KB, Lee K-M, Pascoe B et al. Genome-wide association study of signature genetic alterations among Pseudomonas aeruginosa cystic fibrosis isolates. PLoS Pathog 2021; 17:e1009681 [View Article] [PubMed]
    [Google Scholar]
  7. Bassetti M, Vena A, Croxatto A, Righi E, Guery B. How to manage Pseudomonas aeruginosa infections. DIC 2018; 7:1–18 [View Article]
    [Google Scholar]
  8. Yang F, Liu C, Ji J, Cao W, Ding B et al. Molecular characteristics, antimicrobial resistance, and biofilm formation of Pseudomonas aeruginosa isolated from patients with aural infections in Shanghai, China. Infect Drug Resist 2021; 14:3637–3645 [View Article] [PubMed]
    [Google Scholar]
  9. Wayne P. Clinical and Laboratory Standards Institute (CLSI). In Performance Standards for Antimicrobial Susceptibility Testing vol M100 2021
    [Google Scholar]
  10. Bolger AM, Lohse M, Usadel B. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 2014; 30:2114–2120 [View Article] [PubMed]
    [Google Scholar]
  11. Jackman SD, Vandervalk BP, Mohamadi H, Chu J, Yeo S et al. ABySS 2.0: resource-efficient assembly of large genomes using a Bloom filter. Genome Res 2017; 27:768–777 [View Article] [PubMed]
    [Google Scholar]
  12. Bankevich A, Nurk S, Antipov D, Gurevich AA, Dvorkin M et al. SPAdes: a new genome assembly algorithm and its applications to single-cell sequencing. J Comput Biol 2012; 19:455–477 [View Article] [PubMed]
    [Google Scholar]
  13. Koren S, Walenz BP, Berlin K, Miller JR, Bergman NH et al. Canu: scalable and accurate long-read assembly via adaptive k-mer weighting and repeat separation. Genome Res 2017; 27:722–736 [View Article] [PubMed]
    [Google Scholar]
  14. Seemann T. Prokka: rapid prokaryotic genome annotation. Bioinformatics 2014; 30:2068–2069 [View Article] [PubMed]
    [Google Scholar]
  15. Croucher NJ, Page AJ, Connor TR, Delaney AJ, Keane JA et al. Rapid phylogenetic analysis of large samples of recombinant bacterial whole genome sequences using Gubbins. Nucleic Acids Res 2015; 43:e15 [View Article] [PubMed]
    [Google Scholar]
  16. Stamatakis A. RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies. Bioinformatics 2014; 30:1312–1313 [View Article] [PubMed]
    [Google Scholar]
  17. Bouckaert R, Heled J, Kühnert D, Vaughan T, Wu C-H et al. BEAST 2: a software platform for Bayesian evolutionary analysis. PLoS Comput Biol 2014; 10:e1003537 [View Article] [PubMed]
    [Google Scholar]
  18. Arredondo-Alonso S, Rogers MRC, Braat JC, Verschuuren TD, Top J et al. mlplasmids: a user-friendly tool to predict plasmid- and chromosome-derived sequences for single species. Microb Genom 2018; 4:11 [View Article] [PubMed]
    [Google Scholar]
  19. Li H. Minimap2: pairwise alignment for nucleotide sequences. Bioinformatics 2018; 34:3094–3100 [View Article] [PubMed]
    [Google Scholar]
  20. Yang Z. PAML 4: phylogenetic analysis by maximum likelihood. Mol Biol Evol 2007; 24:1586–1591 [View Article] [PubMed]
    [Google Scholar]
  21. Brynildsrud O, Bohlin J, Scheffer L, Eldholm V. Rapid scoring of genes in microbial pan-genome-wide association studies with Scoary. Genome Biol 2016; 17: [View Article] [PubMed]
    [Google Scholar]
  22. Darch SE, McNally A, Harrison F, Corander J, Barr HL et al. Recombination is a key driver of genomic and phenotypic diversity in a Pseudomonas aeruginosa population during cystic fibrosis infection. Sci Rep 2015; 5:7649 [View Article] [PubMed]
    [Google Scholar]
  23. Lamont IL, Konings AF, Reid DW. Iron acquisition by Pseudomonas aeruginosa in the lungs of patients with cystic fibrosis. Biometals 2009; 22:53–60 [View Article] [PubMed]
    [Google Scholar]
  24. Li Y, Chen L, Zhang P, Bhagirath AY, Duan K. ClpV3 of the H3-type VI secretion system (H3-T6SS) affects multiple virulence factors in Pseudomonas aeruginosa. Front Microbiol 2020; 11:1096 [View Article] [PubMed]
    [Google Scholar]
  25. Parkins MD, Ceri H, Storey DG. Pseudomonas aeruginosa GacA, a factor in multihost virulence, is also essential for biofilm formation. Mol Microbiol 2001; 40:1215–1226 [View Article] [PubMed]
    [Google Scholar]
  26. Lee K, Lee K-M, Go J, Ryu J-C, Ryu J-H et al. The ferrichrome receptor A as a new target for Pseudomonas aeruginosa virulence attenuation. FEMS Microbiol Lett 2016; 363:11 [View Article] [PubMed]
    [Google Scholar]
  27. Magiorakos A-P, Srinivasan A, Carey RB, Carmeli Y, Falagas ME et al. Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clin Microbiol Infect 2012; 18:268–281 [View Article] [PubMed]
    [Google Scholar]
  28. Blunston MA, Yonovitz A, Woodahl EL, Smolensky MH. Gentamicin-induced ototoxicity and nephrotoxicity vary with circadian time of treatment and entail separate mechanisms. Chronobiol Int 2015; 32:1223–1232 [View Article] [PubMed]
    [Google Scholar]
  29. Shaw KJ, Rather PN, Hare RS, Miller GH. Molecular genetics of aminoglycoside resistance genes and familial relationships of the aminoglycoside-modifying enzymes. Microbiol Rev 1993; 57:138–163 [View Article] [PubMed]
    [Google Scholar]
  30. Torres Ortiz A, Coronel J, Vidal JR, Bonilla C, Moore DAJ et al. Genomic signatures of pre-resistance in Mycobacterium tuberculosis. Nat Commun 2021; 12:7312 [View Article] [PubMed]
    [Google Scholar]
  31. Treepong P, Kos VN, Guyeux C, Blanc DS, Bertrand X et al. Global emergence of the widespread Pseudomonas aeruginosa ST235 clone. Clin Microbiol Infect 2018; 24:258–266 [View Article] [PubMed]
    [Google Scholar]
  32. Li XZ, Barré N, Poole K. Influence of the MexA-MexB-oprM multidrug efflux system on expression of the MexC-MexD-oprJ and MexE-MexF-oprN multidrug efflux systems in Pseudomonas aeruginosa. J Antimicrob Chemother 2000; 46:885–893 [View Article] [PubMed]
    [Google Scholar]
  33. Bolard A, Plésiat P, Jeannot K. Mutations in gene fusA1 as a novel mechanism of aminoglycoside resistance in clinical strains of Pseudomonas aeruginosa. Antimicrob Agents Chemother 2018; 62:e01835-17 [View Article] [PubMed]
    [Google Scholar]
  34. Khaledi A, Weimann A, Schniederjans M, Asgari E, Kuo T-H et al. Predicting antimicrobial resistance in Pseudomonas aeruginosa with machine learning-enabled molecular diagnostics. EMBO Mol Med 2020; 12:e10264 [View Article] [PubMed]
    [Google Scholar]
  35. Comolli JC, Hauser AR, Waite L, Whitchurch CB, Mattick JS et al. Pseudomonas aeruginosa gene products PilT and PilU are required for cytotoxicity in vitro and virulence in a mouse model of acute pneumonia. Infect Immun 1999; 67:3625–3630 [View Article] [PubMed]
    [Google Scholar]
  36. Pier GB. Pseudomonas aeruginosa lipopolysaccharide: a major virulence factor, initiator of inflammation and target for effective immunity. Int J Med Microbiol 2007; 297:277–295 [View Article] [PubMed]
    [Google Scholar]
  37. Ozer EA, Nnah E, Didelot X, Whitaker RJ, Hauser AR. The Population Structure of Pseudomonas aeruginosa Is Characterized by Genetic Isolation of exoU+ and exoS+ Lineages. Genome Biol Evol 2019; 11:1780–1796 [View Article] [PubMed]
    [Google Scholar]
  38. Kus JV, Tullis E, Cvitkovitch DG, Burrows LL. Significant differences in type IV pilin allele distribution among Pseudomonas aeruginosa isolates from cystic fibrosis (CF) versus non-CF patients. Microbiology 2004; 150:1315–1326 [View Article] [PubMed]
    [Google Scholar]
  39. Zhang X, Wang L, Li D, Li P, Yuan L et al. An incp-2 plasmid sublineage associated with dissemination of blaimp-45 among carbapenem-resistant Pseudomonas aeruginosa. Emerg Microbes Infect 2021; 10:442–449
    [Google Scholar]
  40. Frasson I, Cavallaro A, Bergo C, Richter SN, Palù G. Prevalence of aac(6’)-Ib-cr plasmid-mediated and chromosome-encoded fluoroquinolone resistance in Enterobacteriaceae in Italy. Gut Pathog 2011; 3:12 [View Article] [PubMed]
    [Google Scholar]
  41. Wang J, Li C, Yang H, Mushegian A, Jin S. A novel serine/threonine protein kinase homologue of Pseudomonas aeruginosa is specifically inducible within the host infection site and is required for full virulence in neutropenic mice. J Bacteriol 1998; 180:6764–6768 [View Article] [PubMed]
    [Google Scholar]
  42. Holm A, Karlsson T, Vikström E. Pseudomonas aeruginosa lasI/rhlI quorum sensing genes promote phagocytosis and aquaporin 9 redistribution to the leading and trailing regions in macrophages. Front Microbiol 2015; 6:915 [View Article] [PubMed]
    [Google Scholar]
  43. Chung The H, Boinett C, Pham Thanh D, Jenkins C, Weill F-X et al. Dissecting the molecular evolution of fluoroquinolone-resistant Shigella sonnei. Nat Commun 2019; 10:4828 [View Article] [PubMed]
    [Google Scholar]
  44. Agnello M, Finkel SE, Wong-Beringer A. Fitness cost of fluoroquinolone resistance in clinical isolates of Pseudomonas aeruginosa differs by type III secretion genotype. Front Microbiol 2016; 7:1591 [View Article] [PubMed]
    [Google Scholar]
  45. Leighton TL, Buensuceso RNC, Howell PL, Burrows LL. Biogenesis of Pseudomonas aeruginosa type IV pili and regulation of their function. Environ Microbiol 2015; 17:4148–4163 [View Article] [PubMed]
    [Google Scholar]
  46. Torrens G, van der Schalk TE, Cortes-Lara S, Timbermont L, Del Barrio-Tofiño E et al. Susceptibility profiles and resistance genomics of Pseudomonas aeruginosa isolates from European ICUs participating in the ASPIRE-ICU trial. J Antimicrob Chemother 2022; 77:1862–1872 [View Article] [PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/mgen/10.1099/mgen.0.000989
Loading
/content/journal/mgen/10.1099/mgen.0.000989
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error