Genetically Diversity of Pseudomonas aeruginosa Isolated from Chronic Suppurative Otitis Media with Respect to Their Antibiotic Sensitivity Pattern

  • Mahesh Chandra Sahu
  • Santosh Kumar Swain
  • Subrat Kumar Kar
Original Article


Pseudomonas aeruginosa is an important chronic suppurative otitis media (CSOM) pathogen that exhibits multiple resistances to antibiotics with increasing frequency, making patient treatment more difficult. The aim of the study is to ascertain the genetically diversity of this clinically isolated P. aeruginosa with inter simple sequence repeat (ISSR) markers. All 25 P. aeruginosa were isolated from CSOM patients by taking their ear swabs and culturing on blood agar and MacConkey agar. All strains were identified with morphological characters and biochemical testing. The antimicrobial susceptibility testing was carried out according to National Committee for Clinical Laboratory Standards. ISSR was used to study the genetic diversity of P. aeruginosa. Clinically CSOM isolated 25 P. aeruginosa were 88% Ciprofloxacin resistant and similarly resistant to other antibiotics were documented. The study has been made using ISSR marker to find out the genomic relation among the strains/populations of P. aeruginosa. The result was shown that maximum similarity (80%) was between S-11 and S-13 and minimum (28.2%) was between S-4 and S-16 with an average similarity of 53.2%. The dendogram showed a distinct separation in between all the strains/populations of P. aeruginosa. The strains/populations were broken up into two main clusters in which small one bear two strains/populations (S-4 and S-9) and another cluster have another 23 strains/populations. These 23 strains were also separated to form subcluster by having different range of small clades. The genetically diversity of pathogenic P. aeruginosa present in CSOM at our hospital indicates the coexistence different strains due to different antibiotic sensitivity patterns. The conventional culture and sensitivity methods are time consuming whereas in PCR, it will detect within 4–6 h for effective antibiotic. Basing upon the banding pattern with ISSR primers, clinicians can prescribe the effective antibiotics accordingly for CSOM patients in the same day.


Pseudomonas aeruginosa CSOM Multi-resistant ISSR Genetical diversity 



This work was supported by the NPDF research project file no PDF/2016/000772 on CSOM, from SERB, DST, Govt. of India, New Delhi.


  1. 1.
    Cripps AW, Kyd J (2003) Bacterial otitis media: current vaccine development strategies. Immunol Cell Biol 81(1):46–51CrossRefPubMedGoogle Scholar
  2. 2.
    Kong K, Coates HLC (2009) Natural history, definitions, risk factors and burden of otitis media. Med J Aust 191(9):s39–s43PubMedGoogle Scholar
  3. 3.
    Monasta L, Ronfani L, Marchetti F, Montico M, Brumatti LV, Bavcar A et al (2012) Burden of disease caused by otitis media: systematic review and global estimates. PLoS ONE 7(4):1–12CrossRefGoogle Scholar
  4. 4.
    Rovers MM (2008) The burden of otitis media. Vaccine 26(suppl 7):G2–G4CrossRefPubMedGoogle Scholar
  5. 5.
    Physicians AA (2004) Otitis media with effusion. Pediatrics 113:1412–1429 (PMID: 15121966) CrossRefGoogle Scholar
  6. 6.
    Massa HM, Cripps AW, Lehmann D (2009) Otitis media: viruses, bacteria, biofilms and vaccines. Med J Aust 191(9):s44–s49PubMedGoogle Scholar
  7. 7.
    Wang EW, Jung JY, Pashia ME, Nason R, Scholnick S, Chole RA (2005) Otopathogenic Pseudomonas aeruginosa strains as competent biofilm formers. Arch Otolaryngol Head Neck Surg 131:983–989CrossRefPubMedGoogle Scholar
  8. 8.
    Sadikot RT, Blackwell TS, Christman JW, Prince AS (2005) Pathogen–host interactions in Pseudomonas aeruginosa pneumonia. Am J Respir Crit Care Med 171:1209–1223CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Mizgerd JP, Horwitz BH, Quillen HC, Scott ML, Doerschuk CM (1999) Effects of CD18 deficiency on the emigration of murine neutrophils during pneumonia. J Immunol 163:995–999PubMedGoogle Scholar
  10. 10.
    Nieuwenhuis EE, Matsumoto T, Exley M et al (2002) CD1 dependent macrophage-mediated clearance of Pseudomonas aeruginosa from lung. Nat Med 8:588–593CrossRefPubMedGoogle Scholar
  11. 11.
    Sader HS, Huband MD, Castanheira M, Flamm RK (2017) Pseudomonas aeruginosa antimicrobial susceptibility results from 4 years (2012–2015) of the international network for optimal resistance monitoring program in the United States. Antimicrobial Agents Chemother 61(3):e02252-16CrossRefGoogle Scholar
  12. 12.
    Yayan J, Ghebremedhin B, Rasche K (2015) Antibiotic resistance of Pseudomonas aeruginosa in pneumonia at a single University Hospital Center in Germany over a 10-year period. PloS ONE 10(10):e0139836CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Forbes BA, Sahm DF, Weissfeld AS (2002) Bailey and Scott’s diagnostics in microbiology. Elsevier, Maryland Heights, pp 1–783Google Scholar
  14. 14.
    Pseudomonas PT, Parker T, Leslie M, Collier H (eds) (1990) Principles of bacteriology, virology and immunology, vol 2, 8th edn. Edward Arnold, New York, pp 255–273Google Scholar
  15. 15.
    Sahu MC, Dubey D, Rath S, Debata NK, Padhy RN (2012) Multidrug resistance of Pseudomonas aeruginosa as known from surveillance of nosocomial and community infections in an Indian teaching hospital. J Publ Health 20(4):413–423CrossRefGoogle Scholar
  16. 16.
    Bauer AM, Kirby WMM, Sherris JC, Turck M (1966) Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol 45:493–496CrossRefPubMedGoogle Scholar
  17. 17.
    Wayne PA (2002) National committee for clinical laboratory standards. Perform Stand Antimicrobial Disc Suscept Test 12:01–53Google Scholar
  18. 18.
    Gherardi M, Mangin B, Goffinet B, Bonnet D, Huguet T (1998) A method to measure genetic distance between allogamous populations of alfalfa (Medicago sativa) using RAPD molecular markers. Theor Appl Genet 96(3–4):406–412CrossRefPubMedGoogle Scholar
  19. 19.
    Real R, Vargas JM (1996) The probabilistic basis of Jaccard’s index of similarity. Syst Biol 45(3):380–385CrossRefGoogle Scholar
  20. 20.
    Kumar S, Tamura K, Nei M (1994) MEGA molecular evolutionary genetics analysis software for microcomputers. Bioinformatics 10(2):189–191CrossRefGoogle Scholar
  21. 21.
    Mantel N (1967) The detection of disease clustering and a generalized regression approach. Cancer Res 27(2 Part 1):209–220PubMedGoogle Scholar
  22. 22.
    Rohlf FJ (1997) NTSYS-pc version 2.0. Exeter Software, Applied Biostatistics IncorporatedGoogle Scholar
  23. 23.
    Prevost A, Wilkinson MJ (1999) A new system of comparing PCR primers applied to ISSR fingerprinting of potato cultivars. Theor Appl Genet 98(1):107–112CrossRefGoogle Scholar
  24. 24.
    Smith JS, Chin EC, Shu H, Smith OS, Wall SJ, Senior ML, Mitchell SE, Kresovich S, Ziegle J (1997) An evaluation of the utility of SSR loci as molecular markers in maize (Zea mays L.): comparisons with data from RFLPs and pedigree. Theor Appl Genet 95(1–2):163–173CrossRefGoogle Scholar
  25. 25.
    Ghislain M, Zhang D, Fajardo D, Huamán Z, Hijmans RJ (1999) Marker-assisted sampling of the cultivated Andean potato Solanum phureja collection using RAPD markers. Genet Resour Crop Evol 46(6):547–555CrossRefGoogle Scholar
  26. 26.
    Tenover FC, Arbeit RD, Goering RV, Mickelsen PA, Murray BE, Persing DH, Swaminathan B (2008) Interpreting chromosomal Mohammad Taheri Z, et al. 39. Tanaffos 7(1):32–39Google Scholar
  27. 27.
    Speijer H, Savelkoul PH, Bonten MJ, Stobberingh EE, Tjhie JH (1999) Application of different genotyping methods for Pseudomonas aeruginosa in a setting of endemicity in an intensive care unit. J Clin Microbiol 37(11):3654–3661PubMedPubMedCentralGoogle Scholar
  28. 28.
    Anthony M, Rose B, Pegler MB, Elkins M, Service H, Thamotharampillai K (2002) Genetic analysis of Pseudomonas aeruginosa isolates from the sputa of Australian adult cystic fibrosis patients. J Clin Microbiol 40(8):2772–2778CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    BAILEY & SCPTT’S. Diagnostic microbiology, 10th ed. MosbyGoogle Scholar
  30. 30.
    Loureiro MM, de Moraes BA, Mendonca VL, Quadra MR, Pinheiro GS, Asensi MD (2002) Pseudomonas aeruginosa: study of antibiotic resistance and molecular typing in hospital infection cases in a neonatal intensive care unit from Rio de Janeiro City, Brazil. Mem Inst Oswaldo Cruz 97(3):387–394CrossRefPubMedGoogle Scholar
  31. 31.
    Renders N, Römling Y, Verbrugh H, van Belkum A (1996) Comparative typing of Pseudomonas aeruginosa by random amplification of polymorphic DNA or pulsed-field gel electrophoresis of DNA macrorestriction fragments. J Clin Microbiol 34(12):3190–3195PubMedPubMedCentralGoogle Scholar
  32. 32.
    Campbell M, Mahenthiralingam E, Speert DP (2000) Evaluation of random amplified polymorphic DNA typing of Pseudomonas aeruginosa. J Clin Microbiol 38(12):4614–4615PubMedPubMedCentralGoogle Scholar
  33. 33.
    Pujana I, Gallego L, Martín G, López F, Canduela J, Cisterna R (1999) Epidemiological analysis of sequential Pseudomonas aeruginosa isolates from chronic bronchiectasis patients without cystic fibrosis. J Clin Microbiol 37(6):2071–2073PubMedPubMedCentralGoogle Scholar
  34. 34.
    Sader HS (2000) Antimicrobial resistance in Brazil: comparison of results from two multicenter studies. Braz J Infect Dis 4(2):91–99PubMedGoogle Scholar
  35. 35.
    Stewart RM, Wiehlmann L, Ashelford KE, Preston SJ, Frimmersdorf E, Campbell BJ, Neal TJ, Hall N, Tuft S, Kaye SB, Winstanley C (2011) Genetic characterization indicates that a specific subpopulation of Pseudomonas aeruginosa is associated with keratitis infections. J Clin Microbiol 49(3):993–1003CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Association of Otolaryngologists of India 2018

Authors and Affiliations

  • Mahesh Chandra Sahu
    • 1
  • Santosh Kumar Swain
    • 2
  • Subrat Kumar Kar
    • 3
  1. 1.Directorate of Medical Research, IMS and SUM HospitalSiksha ‘O’ Anusandhan (Deemed to be University)BhubaneswarIndia
  2. 2.Department of Otorhinolaryngology, IMS and SUM HospitalSiksha ‘O’ Anusandhan (Deemed to be University)BhubaneswarIndia
  3. 3.Department of Microbiology, IMS and SUM HospitalSiksha ‘O’ Anusandhan (Deemed to be University)BhubaneswarIndia

Personalised recommendations