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Isolation, characterization and efficacy of phage MJ2 against biofilm forming multi-drug resistant Enterobacter cloacae

  • Muhsin Jamal
  • Saadia Andleeb
  • Fazal Jalil
  • Muhammad Imran
  • Muhammad Asif Nawaz
  • Tahir Hussain
  • Muhammad Ali
  • Sadeeq ur Rahman
  • Chythanya Rajanna Das
Original Article

Abstract

Biofilm is involved in a variety of infections, playing a critical role in the chronicity of infections. Enterobacter cloacae is a biofilm-forming and multi-drug-resistant (MDR) nosocomial pathogen leading to significant morbidity and mortality. This study aimed at isolation of a bacteriophage against MDR clinical strain of E. cloacae and its efficacy against bacterial planktonic cells and biofilm. A bacteriophage MJ2 was successfully isolated from wastewater and was characterized. The phage exhibited a wide range of thermal and pH stability and demonstrated considerable adsorption to host bacteria in the presence of CaCl2 or MgCl2. Transmission electron microscopy (TEM) showed MJ2 head as approximately 62 and 54 nm width and length, respectively. It had a short non-contractile tail and was characterized as a member of the family Podoviridae [order Caudovirales]. The phage MJ2 was found to possess 11 structural proteins (12–150 kDa) and a double-stranded DNA genome with an approximate size of 40 kb. The log-phase growth of E. cloacae both in biofilm and suspension was significantly reduced by the phage. The E. cloacae biofilm was formed under different conditions to evaluate the efficacy of MJ2 phage. Variable reduction pattern of E. cloacae biofilm was observed while treating it for 4 h with MJ2, i.e., biofilm under static conditions. The renewed media with intervals of 24, 72, and 120 h showed biomass decline of 2.8-, 3-, and 3.5-log, respectively. Whereas, the bacterial biofilm formed with dynamic conditions with refreshing media after 24, 72, and 120 h demonstrated decline in growth at 2.5-, 2.6-, and 3.3-log, respectively. It was, therefore, concluded that phage MJ2 possessed considerable inhibitory effects on MDR E. cloacae both in planktonic and biofilm forms.

Notes

Acknowledgments

K. Kelley from Interdisciplinary Center for Biotechnology Research, UF, USA, helped in TEM studies. Dr. Nighat, Railway General Hospital, Pakistan, provided bacterial strains to establish host range of phage MJ2. Furthermore, anonymous reviewers for their critical comments are highly appreciated.

Funding

The study was sponsored by Higher Education Commission, Pakistan.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Institute of Microbiology, Academy of Sciences of the Czech Republic, v.v.i. 2018

Authors and Affiliations

  • Muhsin Jamal
    • 1
    • 2
    • 3
  • Saadia Andleeb
    • 2
    • 4
  • Fazal Jalil
    • 4
    • 5
  • Muhammad Imran
    • 5
    • 6
  • Muhammad Asif Nawaz
    • 6
    • 7
  • Tahir Hussain
    • 1
  • Muhammad Ali
    • 7
    • 8
  • Sadeeq ur Rahman
    • 3
    • 8
  • Chythanya Rajanna Das
    • 2
    • 3
  1. 1.Department of MicrobiologyAbdul Wali Khan UniversityMardanPakistan
  2. 2.Emerging Pathogens Institute (EPI)University of Florida (UF)GainesvilleUSA
  3. 3.College of Veterinary Sciences & Animal HusbandryAbdul Wali Khan UniversityMardanPakistan
  4. 4.Atta-ur-Rahman School of Applied BiosciencesNational University of Sciences and TechnologyIslamabadPakistan
  5. 5.Department of BiotechnologyAbdul Wali Khan UniversityMardanPakistan
  6. 6.Department of MicrobiologyUniversity of Health SciencesLahorePakistan
  7. 7.Department of BiotechnologyShaheed Benazir Bhutto UniversitySheringalPakistan
  8. 8.Department of Life Sciences, School of ScienceUniversity of Management and TechnologyLahorePakistan

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