Advertisement

Probiotics and Antimicrobial Proteins

, Volume 7, Issue 2, pp 172–180 | Cite as

Protease-Sensitive Inhibitory Activity of Cell-free Supernatant of Lactobacillus crispatus 156 Synergizes with Ciprofloxacin, Moxifloxacin and Streptomycin Against Pseudomonas aeruginosa: An In Vitro Study

  • Sukhraj KaurEmail author
  • Preeti Sharma
Article

Abstract

Ciprofloxacin and streptomycin are frequently prescribed for the treatment of medical conditions originating due to infection by Pseudomonas aeruginosa. However, fluoroquinolone administration has been linked to the outgrowth of Clostridium difficile pathogen, especially in immunocompromised patients. Secondly, frequent administration of antibiotics may lead to development of resistance in the pathogens. Thus, there is a need to explore innovative adjunct therapies to lower the therapeutic doses of the antibiotics. Herein, we evaluated the synergism, if any, between conventional antibiotics and the cell-free supernatant (CFS) of vaginal Lactobacillus crispatus 156 against P. aeruginosa MTCC 741. L. crispatus 156 was isolated from the human vaginal tract, and its CFS had broad-spectrum antimicrobial activity against various Gram-positive and Gram-negative pathogens, including P. aeruginosa. The inhibitory substance present in the CFS completely lost its activity after treatment with proteinases and was resistant to temperatures up to 80 °C and pH ranging from 2 to 6. The cumulative production of the inhibitory substance in CFS was studied, and it showed that the secretion of the inhibitory substance was initiated in middle log phase of growth and peaked in late log phase. Further, CFS synergized the activities of ciprofloxacin, moxifloxacin, and streptomycin as evaluated in terms of checkerboard titrations. It lowered the minimum inhibitory concentration (MIC) of ciprofloxacin by almost 30 times and MIC of both moxifloxacin and streptomycin by 8 times. Interestingly, pepsin treatment of CFS caused the complete abrogation of its synergistic effect with all the three antibiotics. Thus, from the study, it can be concluded that probiotic-based alternative therapeutic regimen can be designed for the treatment of P. aeruginosa infections.

Keywords

Lactobacillus crispatus Probiotic Fluoroquinolones Pseudomonas aeruginosa Synergistic activity 

Notes

Acknowledgments

This work was funded by a research grant to Dr. Sukhraj Kaur by the University Grants Commission (UGC; Vide F.No. 42-478/2013 (SR), dated: 22/03/2013), New Delhi, India. Ms. Preeti Sharma is grateful to UGC for the award of Junior Research Fellowship under the scheme University of Potential for Excellence, granted to Guru Nanak Dev University, Amritsar, India.

Conflict of Interest

Sukhraj Kaur and Preeti Sharma declares that they have no conflict of interest.

References

  1. 1.
    National Nosocomial Infections Surveillance system report (1999) Data summary from January 1990–May 1999. Am J Infect Control 2:520–532Google Scholar
  2. 2.
    Goossens H (2003) Susceptibility of multi-drug-resistant Pseudomonas aeruginosa in intensive care units: results from the European MYSTIC study group. Clin Microbiol Infect 9:980–983CrossRefGoogle Scholar
  3. 3.
    Poole K (2004) Efflux-mediated multi resistance in Gram-negative bacteria. Clin Microbiol Infect 10:12–26CrossRefGoogle Scholar
  4. 4.
    National Nosocomial Infections Surveillance system report (2004) Data summary from January 1992 through June 2004. Am J Infect Control 32:470–485CrossRefGoogle Scholar
  5. 5.
    Pépin J, Saheb N, Coulombe M, Alary M, Corriveau M, Authier S et al (2005) Emergence of fluoroquinolones as the predominant risk factor for Clostridium difficile–associated diarrhea: a cohort study during an epidemic in Quebec. Clin Infect Dis 41:1254–1260CrossRefGoogle Scholar
  6. 6.
    Morita Y, Tomida J, Kawamura Y (2014) Responses of Pseudomonas aeruginosa to antimicrobials. Front Microbiol. doi: 10.3389/fmicb.2013.00422 Google Scholar
  7. 7.
    Forestier C, Guelon D, Cluytens V, Gillart T, Sirot J, de Champs C (2008) Oral probiotic and prevention of Pseudomonas aeruginosa infections: a randomized, double-blind, placebo-controlled pilot study in ICU-patients. Crit Care 12:R69. doi: 10.1186/cc6907 CrossRefGoogle Scholar
  8. 8.
    Stapleton AE, Au-Yeung M, Hooton TM, Fredericks DN, Roberts PL, Czaja CA et al (2011) Randomized, placebo-controlled phase 2 clinical trial of a Lactobacillus crispatus probiotic given intravaginally for prevention of recurrent urinary tract infection. Clin Infect Dis 52:1212–1217CrossRefGoogle Scholar
  9. 9.
    Pavlova SI, Kilic AO, Kilic SS, So JS, Nader-Macias ME, Simoes JA et al (2002) Genetic diversity of vaginal lactobacilli from women in different countries based on 16S rRNA gene sequences. J Appl Microbiol 92:451–459CrossRefGoogle Scholar
  10. 10.
    Lakshminarayanan B, Guinane CM, O’Connor PM, Coakley M, Hill C, Stanton C et al (2013) Isolation and characterization of bacteriocin-producing bacteria from the intestinal microbiota of elderly Irish subjects. J Appl Microbiol 114:886–898CrossRefGoogle Scholar
  11. 11.
    Kim J, Rajagopal SN (2001) Antibacterial activities of Lactobacillus crispatus ATCC 33820 and Lactobacillus gasseri ATCC 33323. J Microbiol 39:146–148Google Scholar
  12. 12.
    Tahara T, Kanatani K (1997) Isolation and partial characterisation of crispacin, a cell-associated bacteriocin produced by Lactobacillus crispatus JCM 2009. FEMS Microbiol Lett 147:287–290CrossRefGoogle Scholar
  13. 13.
    de Man JC, Rogosa M, Sharpe ME (1960) A medium for the cultivation of lactobacilli. J Appl Bacteriol 23:130–135CrossRefGoogle Scholar
  14. 14.
    Holt JG, Krieg NR, Sneath PH, Staley JT, Williams ST (1994) Bergeys manual of determinative bacteriology, 9th edn. Lippincott Williams and Wilkins, BaltimoreGoogle Scholar
  15. 15.
    Moore E, Arnscheidt A, Kuger A, Strompl C, Mau M (2004) Simplified protocols for the preparation of genomic DNA from bacterial cultures. In: Kowalchuk GA, de Bruijn FJ, Head IM, Van der Zijpp AJ, van Elsas JD (eds) Molecular microbial ecology manual, 2nd edn. Kluwer Academic Publishers, Netherland, pp 3–18Google Scholar
  16. 16.
    Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410CrossRefGoogle Scholar
  17. 17.
    Geis A, Singh J, Teuber M (1983) Potential of lactic streptococci to produce bacteriocin. Appl Environ Microbiol 45:205–211Google Scholar
  18. 18.
    Melancon D, Grenier D (2003) Production and properties of bacteriocin-like inhibitory substances from the swine pathogens Streptococcus suis serotype 2. Appl Environ Microbiol 69:4482–4488CrossRefGoogle Scholar
  19. 19.
    Bauer AW, Kirby MM, Sherris JC, Turck M (1966) Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol 45:493–496Google Scholar
  20. 20.
    National Committee for clinical laboratory standards (2000) Performance standards for antimicrobial disk susceptibility tests, 7th edn. Approved standard M2-A7. NCCLS, WayneGoogle Scholar
  21. 21.
    Ruíz FO, Gerbaldo G, García MJ, Giordano W, Pascual L, Barberis IL (2012) Synergistic effect between two bacteriocin-like inhibitory substances produced by lactobacilli strains with inhibitory activity for Streptococcus agalactiae. Curr Microbiol 64:349–356CrossRefGoogle Scholar
  22. 22.
    Petersen P, Labthavikul P, Jones C, Bradford P (2006) In vitro antibacterial activities of tigecycline in combination with other antimicrobial agents determined by chequerboard and time-kill kinetic analysis. J Antimicrob Chemother 57:573–576CrossRefGoogle Scholar
  23. 23.
    Dicks LMT, Heunis TDJ, Staden DAV, Brand A, Noll KS, Chikindas ML (2011) Medical and personal care applications of bacteriocins produced by Lactic acid bacteria. In: Drider D, Rebuffat S (eds) Prokaryotic antimicrobial peptides: from genes to applications. Springer, New York, pp 391–421CrossRefGoogle Scholar
  24. 24.
    Cotter PD, Ross RP, Hill C (2013) Bacteriocins—a viable alternative to antibiotics? Nat Rev Microbiol 11:95–105CrossRefGoogle Scholar
  25. 25.
    Dobson A, Cotter R, Ross P, Hill C (2012) Bacteriocin production: a probiotic trait? Appl Environ Microbiol 78:1–6. doi: 10.1128/AEM.05576-11 CrossRefGoogle Scholar
  26. 26.
    Al-Mathkhury HJF, Ali AS, Ghafil JA (2011) Antagonistic effect of bacteriocin against urinary catheter associated Pseudomonas aeruginosa biofilm. N Am J Med Sci 3(8):367–370. doi: 10.4297/najms.2011.3367 CrossRefGoogle Scholar
  27. 27.
    Majhenic AC, Matijasic BB, Rogelj I (2003) Chromosomal location of the genetic determinants for bacteriocins produced by Lactobacillus gasseri K7. J Dairy Res 70:199–203. doi: 10.1017/S0022029903006162 CrossRefGoogle Scholar
  28. 28.
    Stover CK, Pham XQ, Erwin AL et al (2000) Complete genome sequence of Pseudomonas aeruginosa PAO1, an opportunistic pathogen. Nature 406:959–964CrossRefGoogle Scholar
  29. 29.
    Gálvez A, Abriouel H, López RL, Omar NB (2007) Bacteriocin-based strategies for food biopreservation. Int J Food Microbiol 120:51–70CrossRefGoogle Scholar
  30. 30.
    Rishi P, Preet S, Kaur P (2011) Effect of L. plantarum cell-free extract and co-trimoxazole against Salmonella typhimurium: a possible adjunct therapy. Ann Clin Microbiol Antimicrob 10:9. doi: 10.1186/1476-0711-10-9 CrossRefGoogle Scholar
  31. 31.
    Giacometti A, Cirioni O, Barcheisi M, Fortuna M, Scarleise E (1999) In vitro activity of cationic peptides alone and in combination with clinically used antimicrobial agents against Pseudomonas aeruginosa. J Antimicrob Chemother 44:641–645CrossRefGoogle Scholar
  32. 32.
    Mataraci E, Dosler S (2012) In vitro activities of antibiotics and antimicrobial cationic peptides alone and in combination against methicillin-resistant Staphylococcus aureus biofilms. Antimicrob Agents Chemother 56:6366–6371CrossRefGoogle Scholar
  33. 33.
    Draper LA, Cotter PD, Hill C, Ross RP (2013) The two peptide lantibiotic lacticin 3147 acts synergistically with polymyxin to inhibit Gram negative bacteria. BMC Microbiol 13:212. doi: 10.1186/1471-2180-13-212 CrossRefGoogle Scholar
  34. 34.
    Drlica K, Zhao XK (1997) DNA gyrase, topoisomerase IV, and the 4-quinolones. Microbiol Mol Biol Rev 61:377–392Google Scholar
  35. 35.
    Hancock RE (1997) Peptide antibiotics. Lancet 349:418–422CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Department of MicrobiologyGuru Nanak Dev UniversityAmritsarIndia

Personalised recommendations