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Highly synergistic activity of melittin with imipenem and colistin in biofilm inhibition against multidrug-resistant strong biofilm producer strains of Acinetobacter baumannii

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Abstract

The rapid increase of drug resistance and failure of available antibiotics to treat biofilm-associated infections is of great health concern. Accordingly, our study aimed to evaluate the synergistic antibacterial, biofilm inhibitory, and biofilm removal activities of melittin in combination with colistin, imipenem, and ciprofloxacin against multidrug-resistant (MDR) strong biofilm producer Acinetobacter baumannii isolates. The kinetics of biofilm formation were evaluated for the isolates for 144 h. Minimum inhibitory concentrations (MICs), minimum bactericidal concentrations (MBCs), minimum biofilm inhibitory concentrations (MBICs), and biofilm removal activities for melittin and combinations with antibiotics were determined. Inhibition of biofilm-associated protein (bap) expression by melittin was evaluated with real-time polymerase chain reaction (PCR). Field emission scanning electron microscopy (FE-SEM) was used to visualize the effect of synergism on the inhibition of biofilm production. The geometric means of the fractional inhibitory concentration index (FICi) for melittin–colistin, melittin–imipenem, and melittin–ciprofloxacin combinations were calculated as 0.31, 0.24, and 0.94, respectively. Comparing the geometric means of the removal activity for melittin, colistin, imipenem, and combinations of them in both 6 and 24 h showed a significant difference between the groups (p-value < 0.05). Exposure to melittin induced a statistically significant downregulation of bap mRNA levels in all isolates at sub-MIC doses. Analysis of the FE-SEM results demonstrated that the synergism of melittin–colistin at 0.125–0.25 μg inhibited biofilm formation completely. In conclusion, our findings indicate that melittin possesses considerable potential for use in combination with colistin and imipenem to treat infections caused by MDR strong biofilm producer A. baumannii isolates.

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Abbreviations

AMP:

Antimicrobial peptides

MDR:

Multidrug-resistant

bap:

Biofilm-associated protein

CFU:

Colony-forming units

Col:

Colistin

Cip:

Ciprofloxacin

FE-SEM:

Field emission scanning electron microscopy

FICi:

Fractional inhibitory concentration index

Imp:

Imipenem

IBB:

Interbacterial biofilm

MHB:

Mueller-Hinton broth

MIC:

Minimum inhibitory concentration

MBC:

Minimum bactericidal concentration

Mel:

Melittin

MBIC:

Minimum biofilm inhibitory concentration

SBBs:

Small biofilm biomasses

TSB:

Tryptic soy broth

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Acknowledgements

This investigation is a part of the Ph.D. thesis approved and financially supported by the Vice-Chancellor of Research and Technology of Hamadan University of Medical Sciences, Hamadan, Iran and technically supported by Pasteur Institute of Iran, Tehran, Iran.

Author information

Authors and Affiliations

  1. Department of Microbiology, Faculty of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran

    Ali Mohammadi Bardbari, Mohammad Reza Arabestani & Mohammad Yousef Alikhani

  2. Department of Epidemiology, School of Public Health, Hamadan University of Medical Sciences, Hamadan, Iran

    Manoochehr Karami

  3. Department of Infectious Diseases, Faculty of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran

    Fariba Keramat

  4. Brucellosis Research Center, Hamadan University of Medical Sciences, Hamadan, Iran

    Fariba Keramat & Mohammad Yousef Alikhani

  5. Venom and Biotherapeutics Molecules Lab., Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran

    Hossein Aghazadeh & Kamran Pooshang Bagheri

Authors
  1. Ali Mohammadi Bardbari
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  2. Mohammad Reza Arabestani
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  3. Manoochehr Karami
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  4. Fariba Keramat
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  5. Hossein Aghazadeh
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  6. Mohammad Yousef Alikhani
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  7. Kamran Pooshang Bagheri
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Contributions

A.M.B. performed all experiments and analysis and also contributed to writing the manuscript. M.R.A., M.K., and F.K. served as advisors. H.A. contributed to the FE-SEM experiments. M.Y.A. contributed as a supervisor and also revision of the manuscript. K.P.B. suggested the implementation of melittin, contributed as a supervisor, and also in the writing, revision, and redaction of the manuscript.

Corresponding authors

Correspondence to Mohammad Yousef Alikhani or Kamran Pooshang Bagheri.

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None of the authors have any conflicts of interest related to this article.

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Bardbari, A.M., Arabestani, M.R., Karami, M. et al. Highly synergistic activity of melittin with imipenem and colistin in biofilm inhibition against multidrug-resistant strong biofilm producer strains of Acinetobacter baumannii. Eur J Clin Microbiol Infect Dis 37, 443–454 (2018). https://doi.org/10.1007/s10096-018-3189-7

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  • DOI: https://doi.org/10.1007/s10096-018-3189-7

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