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

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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.

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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.

Author information

Authors and Affiliations

  1. Department of Microbiology, Abdul Wali Khan University, Garden Campus, Mardan, 23200, Pakistan

    Muhsin Jamal & Tahir Hussain

  2. Emerging Pathogens Institute (EPI), University of Florida (UF), Gainesville, FL, USA

    Muhsin Jamal, Saadia Andleeb & Chythanya Rajanna Das

  3. College of Veterinary Sciences & Animal Husbandry, Abdul Wali Khan University, Mardan, Pakistan

    Muhsin Jamal, Sadeeq ur Rahman & Chythanya Rajanna Das

  4. Atta-ur-Rahman School of Applied Biosciences, National University of Sciences and Technology, Islamabad, Pakistan

    Saadia Andleeb & Fazal Jalil

  5. Department of Biotechnology, Abdul Wali Khan University, Mardan, 23200, Pakistan

    Fazal Jalil & Muhammad Imran

  6. Department of Microbiology, University of Health Sciences, Lahore, Pakistan

    Muhammad Imran & Muhammad Asif Nawaz

  7. Department of Biotechnology, Shaheed Benazir Bhutto University, Sheringal, Dir (Upper), Pakistan

    Muhammad Asif Nawaz & Muhammad Ali

  8. Department of Life Sciences, School of Science, University of Management and Technology, C-II Johar Town, Lahore, 54770, Pakistan

    Muhammad Ali & Sadeeq ur Rahman

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  1. Muhsin Jamal
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Correspondence to Muhsin Jamal.

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Jamal, M., Andleeb, S., Jalil, F. et al. Isolation, characterization and efficacy of phage MJ2 against biofilm forming multi-drug resistant Enterobacter cloacae. Folia Microbiol 64, 101–111 (2019). https://doi.org/10.1007/s12223-018-0636-x

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