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Abstract

To isolate specific bacteria from samples constituting the microbiota, it is essential to employ selective media that suppress the growth of resident bacteria other than specific target bacteria. Selective media for clinically important (including , which was previously taxonomically classified as part of the genus ) have been limited because they have been designed for a limited range of species within the genus and require ingredients which are difficult to prepare and handle. This study aimed to develop a selective medium [referred to as and Selective Medium (ASSM)] for the isolation of a broad range of and species from samples mixed with resident bacteria. The composition of ASSM includes yeast extract, agar, brain heart infusion (BHI), levofloxacin (LVFX), fosfomycin (FOM), colistin (CL) and metronidazole (MNZ). Evaluation of the medium using 24 swab samples serially collected from the roots of the teeth of a healthy individual for whom metagenome sequencing data of a saliva sample are publicly available revealed that ASSM adjusted to concentrations of LVFX 0.5 mg l, FOM 5 mg l, CL 1 mg l and MNZ 2 mg l and cultured anaerobically at 35 °C for 7 days enabled the isolation of species from 37.5 % of the samples. The inclusion of CL and MNZ in ASSM can also be useful for samples harbouring other bacterial species. The selective isolation medium is expected to contribute to studies investigating the relationship between these bacteria and their pathogenesis or disease.

Funding
This study was supported by the:
  • “Grants-in-Aid for Scientific Research” from the Ministry of Education, Culture, Sports and Science and and Technology (MEXT) (Award 19J40070 and 21K10129)
    • Principle Award Recipient: HirokoYahara
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License.
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/content/journal/acmi/10.1099/acmi.0.000768.v3
2024-05-10
2024-05-23
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References

  1. Könönen E, Wade WG. Actinomyces and related organisms in human infections. Clin Microbiol Rev 2015; 28:419–442 [View Article] [PubMed]
    [Google Scholar]
  2. Yahara H, Hiraki A, Maruoka Y, Hirabayashi A, Suzuki M et al. Shotgun metagenome sequencing identification of a set of genes encoded by Actinomyces associated with medication-related osteonecrosis of the jaw. PLoS One 2020; 15:e0241676 [View Article] [PubMed]
    [Google Scholar]
  3. Cerrato A, Zanette G, Boccuto M, Angelini A, Valente M et al. Actinomyces and MRONJ: a retrospective study and a literature review. J Stomatol Oral Maxillofac Surg 2021; 122:499–504 [View Article] [PubMed]
    [Google Scholar]
  4. Kövér Z, Bán Á, Gajdács M, Polgár B, Urbán E. Role of Actinomyces spp. and related organisms in the development of medication-related osteonecrosis of the jaw (MRONJ): clinical evidence based on a case series. Eur J Microbiol Immunol 2023; 13:125–134 [View Article] [PubMed]
    [Google Scholar]
  5. Nouioui I, Carro L, García-López M, Meier-Kolthoff JP, Woyke T et al. Genome-based taxonomic classification of the phylum actinobacteria. Front Microbiol 2018; 9:2007 [View Article] [PubMed]
    [Google Scholar]
  6. Zylber LJ, Jordan HV. Development of a selective medium for detection and enumeration of Actinomyces viscosus and Actinomyces naeslundii in dental plaque. J Clin Microbiol 1982; 15:253–259 [View Article] [PubMed]
    [Google Scholar]
  7. Tsuzukibashi O, Fukatsu A, Fuchigami M, Uchibori S, Komine C et al. Isolation and identification methods for Actinomyces israelii involved in actinomycosis. Open J Stomatol 2022; 12:108–118 [View Article]
    [Google Scholar]
  8. Lewis R, McKenzie D, Bagg J, Dickie A. Experience with a novel selective medium for isolation of Actinomyces spp. from medical and dental specimens. J Clin Microbiol 1995; 33:1613–1616 [View Article] [PubMed]
    [Google Scholar]
  9. Ellen RP, Balcerzak-Raczkowski IB. Differential medium for detecting dental plaque bacteria resembling Actinomyces viscosus and Actinomyces naeslundii. J Clin Microbiol 1975; 2:305–310 [View Article] [PubMed]
    [Google Scholar]
  10. Kornman KS, Loesche WJ. New medium for isolation of Actinomyces viscosus and Actinomyces naeslundii from dental plaque. J Clin Microbiol 1978; 7:514–518 [View Article] [PubMed]
    [Google Scholar]
  11. Beighton D, Colman G. A medium for the isolation and enumeration of oral actinomycetaceae from dental plaque. J Dent Res 1976; 55:875–878 [View Article] [PubMed]
    [Google Scholar]
  12. Yahara K, Suzuki M, Hirabayashi A, Suda W, Hattori M et al. Long-read metagenomics using PromethION uncovers oral bacteriophages and their interaction with host bacteria. Nat Commun 2021; 12:27 [View Article] [PubMed]
    [Google Scholar]
  13. Rams TE, Feik D, Mortensen JE, Degener JE, van Winkelhoff AJ. Antibiotic susceptibility of periodontal Enterococcus faecalis. J Periodontol 2013; 84:1026–1033 [View Article] [PubMed]
    [Google Scholar]
  14. Seo Y, Lee G. Antimicrobial resistance pattern in Enterococcus faecalis strains isolated from expressed prostatic secretions of patients with chronic bacterial prostatitis. Korean J Urol 2013; 54:477–481 [View Article] [PubMed]
    [Google Scholar]
  15. Zhanel GG, Zhanel MA, Karlowsky JA. Intravenous fosfomycin: an assessment of its potential for use in the treatment of systemic infections in Canada. Can J Infect Dis Med Microbiol 2018; 2018:8912039 [View Article] [PubMed]
    [Google Scholar]
  16. Steininger C, Willinger B. Resistance patterns in clinical isolates of pathogenic Actinomyces species. J Antimicrob Chemother 2016; 71:422–427 [View Article] [PubMed]
    [Google Scholar]
  17. Smith AJ, Hall V, Thakker B, Gemmell CG. Antimicrobial susceptibility testing of Actinomyces species with 12 antimicrobial agents. J Antimicrob Chemother 2005; 56:407–409 [View Article] [PubMed]
    [Google Scholar]
  18. Takesue Y, Kusachi S, Mikamo H, Sato J, Watanabe A et al. Antimicrobial susceptibility of pathogens isolated from surgical site infections in Japan: comparison of data from nationwide surveillance studies conducted in 2010 and 2014-2015. J Infect Chemother 2017; 23:339–348 [View Article] [PubMed]
    [Google Scholar]
  19. Kastoris AC, Rafailidis PI, Vouloumanou EK, Gkegkes ID, Falagas ME. Synergy of fosfomycin with other antibiotics for Gram-positive and Gram-negative bacteria. Eur J Clin Pharmacol 2010; 66:359–368 [View Article] [PubMed]
    [Google Scholar]
  20. Fonnes S, Holzknecht BJ, Gasbjerg LS, Weisser JJ, Hallberg HW et al. The combination of Fosfomycin, Metronidazole, and recombinant human Granulocyte-Macrophage colony-stimulating factor is stable in vitro and has maintained antibacterial activity. Drug Res 2018; 68:349–354 [View Article] [PubMed]
    [Google Scholar]
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