Journal of Medical Microbiology Papers in Press. Published February 13, 2015 as doi:10.1099/jmm.0.000040
Journal of Medical Microbiology Characterization of Siphoviridae phage Z and studying its efficacy against multi-drug resistant (MDR) Klebsiella pneumoniae planktonic cells and biofilm --Manuscript Draft-Manuscript Number:
Characterization of Siphoviridae phage Z and studying its efficacy against multi-drug resistant (MDR) Klebsiella pneumoniae planktonic cells and biofilm
Phage Z against multi-drug resistant (MDR) Klebsiella pneumoniae
Muhsin Jamal Tahir Hussain Chythanya Rajanna Das Saadia Andleeb
Biofilm is involved in many serious consequences for public health and is a major virulence factor contributing to the chronicity of infections. The aim of the current study was to isolate and characterize a bacteriophage that inhibit multi-drug resistant Klebsiella pneumonia (M) in palnktonic form as well as biofilm. This phage designated as bacteriophage Z was isolated from waste water. Its adsorption rate to its host bacterium was significantly enhanced by MgCl2 and CaCl2. It has a wide range of pH and heat stability. From its one step growth, latent time and burst size was determined that were 24 min and about 320 virions per cell, respectively. As analyzed by transmission electron microscopy, phage Z had a head of width (76±10 nm) and length of (92±14 nm) with an icosahedrons' sides of 38 nm with a non- contractile (200±15 nm) long and (14-29 nm) wide tail belonging to family Siphoviridae of order Caudovirales. Six structural proteins ranging from 18 to 65 kDa were revealed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Its genome was found to be comprised of double stranded DNA with approximate size of 36 kb. Bacteria were grown in suspension and as biofilms to compare the susceptibility of both phenotypes to the phage lytic action. Phage Z was effective in reducing biofilm biomass after 24 h and 48 h and showed more than 2-fold and 3-fold reductions, respectively. Biofilm cells and stationary phase planktonic bacteria were killed at a lower rate than the log-phase planktonic bacteria.
Powered by Editorial Manager® and ProduXion Manager® from Aries Systems Corporation
Manuscript Including References (Word document) Click here to download Manuscript Including References (Word document): Paper K.P (M) Final Revision 2.docx
Characterization of Siphoviridae phage Z and studying its efficacy against multi-drug
resistant (MDR) Klebsiella pneumoniae planktonic cells and biofilm
3 4 5
Phage Z against multi-drug resistant (MDR) Klebsiella pneumoniae
Ackermann, H. and Dubow, M. (1987). Viruses of prokaryotes: general properties of bacteriophages. Boca Raton, Fla. CRC Press, Inc. Adams, M. H. (1959). Bacteriophages. New York, Interscience Publishers, Inc. Archibald, L., Phillips, L., Monnet, D., Mcgowan, J. E., Tenover, F. and Gaynes, R. (1997). Antimicrobial resistance in isolates from inpatients and outpatients in the United States: increasing importance of the intensive care unit. Clin Infect Dis 24, 211-215. Ashelford, K. E., Norris, S. J., Fry, J. C., Bailey, M. J. and Day, M. J. (2000). Seasonal population dynamics and interactions of competing bacteriophages and their host in the rhizosphere. Appl Environ Microbiol 66, 4193-4199. Briandet, R., Lacroix-Gueu, P., Renault, M., Lecart, S., Meylheuc, T., Bidnenko, E., Steenkeste, K., Bellon-Fontaine, M.-N. and Fontaine-Aupart, M.-P. (2008). Fluorescence correlation spectroscopy to study diffusion and reaction of bacteriophages inside biofilms. Appl Environ Microbiol 74, 2135-2143. Capra, M., Quiberoni, A. and Reinheimer, J. (2006). Phages of Lactobacillus casei/paracasei: response to environmental factors and interaction with collection and commercial strains. J Appl Microbiol 100, 334-342. Carson, L., Gorman, S. P. and Gilmore, B. F. (2010). The use of lytic bacteriophages in the prevention and eradication of biofilms of Proteus mirabilis and Escherichia coli. FEMS Immunol Med Microbiol 59, 447-455. Cerca, N., Oliveira, R. and Azeredo, J. (2007). Susceptibility of Staphylococcus epidermidis planktonic cells and biofilms to the lytic action of staphylococcus bacteriophage K. Lett Appl Microbiol 45, 313-317. Corbin, B. D., Mclean, R. J. and Aron, G. M. (2001). Bacteriophage T4 multiplication in a glucose-limited Escherichia coli biofilm. Canadian J Microbiol 47, 680-684. Costerton, J. (1995). Overview of microbial biofilms. J Indust Microbiol 15, 137-140. Costerton, J., Stewart, P. S. and Greenberg, E. (1999). Bacterial biofilms: a common cause of persistent infections. Sci 284, 1318-1322. Costerton, J. W., Geesey, G. and Cheng, K. (1978). How bacteria stick. Scientific American, 238 (1): 86.
Donlan, R. M. (2005). New approaches for the characterization of prosthetic joint biofilms. Clin Orthopaed Related Res 437, 12-19. Donlan, R. M. and Costerton, J. W. (2002). Biofilms: survival mechanisms of clinically relevant microorganisms. Clin Microbiol Rev 15, 167-193. Gilbert, P., Das, J. and Foley, I. (1997). Biofilm susceptibility to antimicrobials. Advances in Dental Res 11, 160-167. Guttman, B., Raya, R. and Kutter, E. (2005). Basic phage biology. Bacteriophages: Biol Applications 4. Haq, I. U., Chaudhry, W. N., Andleeb, S. and Qadri, I. (2012). Isolation and Partial Characterization of a Virulent Bacteriophage IHQ1 Specific for Aeromonas punctata from Stream Water. Microbial Ecol 63, 954-963. Hazem, A. (2002). Effects of temperatures, pH-values, ultra-violet light, ethanol and chloroform on the growth of isolated thermophilic Bacillus phages. The New Microbiologica 25, 469-476. Hughes, K. A., Sutherland, I. W. and Jones, M. V. (1998). Biofilm susceptibility to bacteriophage attack: the role of phage-borne polysaccharide depolymerase. Microbiol 144, 3039-3047. Jamalludeen, N., Johnson, R. P., Friendship, R., Kropinski, A. M., Lingohr, E. J. and Gyles, C. L. (2007). Isolation and characterization of nine bacteriophages that lyse O149 enterotoxigenic Escherichia coli. Vet Microbiol 124, 47-57. Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-685. Mah, T.-F. C. and O'toole, G. A. (2001). Mechanisms of biofilm resistance to antimicrobial agents. Trends in Microbiol 9, 34-39. Monroe, D. (2007). Looking for chinks in the armor of bacterial biofilms. PLoS Biol 5, e307. Moons, P., Werckx, W., Van Houdt, R., Aertsen, A. and Michiels, C. W. (2006). Resistance development of bacterial biofilms against bacteriophage attack. Commun Agric Appl Biol Sci 71, 297-300. Nakai, T. and Park, S. C. (2002). Bacteriophage therapy of infectious diseases in aquaculture. Res Microbiol 153, 13-18. Piracha, Z., Saeed, U., Khurshid, A. and Chaudhary, W. N. (2014). Isolation and Partial Characterization of Virulent Phage Specific against Pseudomonas Aeruginosa. Global J Med Res 14. Podschun, R. and Ullmann, U. (1998). Klebsiella spp. as nosocomial pathogens: epidemiology, taxonomy, typing methods, and pathogenicity factors. Clin Microbiol Rev 11, 589-603. Potera, C. (1999). Forging a link between biofilms and disease. Sci 283, 1837-1839. Rao, V., Ghei, R. and Chambers, Y. (2005). Biofilms research-implications to biosafety and public health. Appl Biosafety 10, 83. Reese, J. F., Dimitracopoulos, G. and Bartell, P. F. (1974). Factors influencing the adsorption of bacteriophage 2 to cells of Pseudomonas aeruginosa. J Virol 13, 22-27. Roy, B., Ackermann, H., Pandian, S., Picard, G. and Goulet, J. (1993). Biological inactivation of adhering Listeria monocytogenes by listeriaphages and a quaternary ammonium compound. Appl Environ Microbiol 59, 2914-2917. Russel, M., Whirlow, H., Sun, T. and Webster, R. (1988). Low-frequency infection of F-bacteria by transducing particles of filamentous bacteriophages. J Bacteriol 170, 5312-5316. Sambrook, J., Fritsch, E. F. and Maniatis, T. (1989). Molecular cloning, Cold spring harbor laboratory press New York. Sillankorva, S., Oliveira, R., Vieira, M. J., Sutherland, I. and Azeredo, J. (2004). Bacteriophage Φ S1 infection of Pseudomonas fluorescens planktonic cells versus biofilms. Biofouling 20, 133-138. Sulakvelidze, A., Alavidze, Z. and Morris, J. G. (2001). Bacteriophage therapy. Antimicrobial Agents and Chemother 45, 649-659. Tenover, F. C. (2001). Development and spread of bacterial resistance to antimicrobial agents: an overview. Clin Infect Dis 33, S108-S115. 19
Van Regenmortel, M. H., Fauquet, C. M., Bishop, D. H., Carstens, E., Estes, M., Lemon, S., Maniloff, J., Mayo, M., Mcgeoch, D. and Pringle, C. (2000). Virus taxonomy: classification and nomenclature of viruses. Seventh report of the International Committee on Taxonomy of Viruses, Academic Press. Vinh, D. C. and Embil, J. M. (2005). Device-related infections: a review. J Long-Term Effects of Med Implants 15. Watanabe, K. and Takesue, S. (1972). The requirement for calcium in infection with Lactobacillus phage. J Gen Viro 17, 19-30. Watnick, P. and Kolter, R. (2000). Biofilm, city of microbes. J Bacteriol 182, 2675-2679. Yang, H., Liang, L., Lin, S. and Jia, S. (2010). Isolation and characterization of a virulent bacteriophage AB1 of Acinetobacter baumannii. BMC Microbiol 10, 131. Zimmer, M., Scherer, S. and Loessner, M. J. (2002). Genomic analysis of Clostridium perfringens bacteriophage φ3626, which integrates into guaA and possibly affects sporulation. J Bacteriol 184, 43594368.
Background: The aim of this study was to evaluate the susceptibility of catheter-related Klebsiella pneumoniae isolates to two biocides (benzalkonium chloride and Deconex) under biofilm and planktonic conditions. Methods: A total of 85 strains of K.
Free-living amoebae (FLA) in aqueous systems are a problem for water network managers and health authorities because some are pathogenic, such as Naegleria fowleri, and they have also been reported to operate as reservoirs and vectors of several path
Klebsiella pneumoniae is among the most frequently recovered etiologic agents from nosocomial infections. This opportunistic pathogen can generate a thick layer of biofilm as one of its important virulence factors, enabling the bacteria to attach to
The treatment of Klebsiella pneumoniae, particularly extended-spectrum β-lactamase (ESBL)-producing K. pneumoniae, is currently a great challenge. Photodynamic antimicrobial chemotherapy is a promising approach for killing antibiotic-resistant bacter
Many bacterial species use their intercellular signaling mechanism called quorum sensing (QS), which is found to be implicated in various factors including bacterial pathogenicity and food spoilage. Interrupting the bacterial communication is an attr
Pseudomonas aeruginosa is one of the Multi-Drug-Resistant organisms most frequently isolated worldwide and, because of a shortage of new antibiotics, bacteriophages are considered an alternative for its treatment. Previously, P. aeruginosa phages wer
The Gram-negative opportunistic pathogen, Klebsiella pneumoniae, is responsible for causing a spectrum of community-acquired and nosocomial infections and typically infects patients with indwelling medical devices, especially urinary catheters, on wh
ϕRS138, a bacteriophage of the family Siphoviridae that lyses Ralstonia solanacearum, was isolated. The genomic DNA of ϕRS138 was 41,941 bp long with a GC content of 65.1 % and contained 56 putative open reading frames. The ϕRS138 genome could be div
Streptococcus equi ssp. zooepidemicus (SEZ) is responsible for a wide variety of infections in many species, including pigs, horses and humans. Biofilm formation is essential for pathogenesis, and the ability to resist antibiotic treatment results in