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Isolation of Bacteriophage and Assessment of its Activity against Biofilms of Uropathogenic Escherichia coli in Jimma Town, South Western Ethiopia

Israel Gudina, Zeleke Gizachew, Delelegn Woyessa, Tesfaye K Tefera

Abstract


Escherichia coli is one of the most commonly associated bacteria with urinary tract infections (UTIs) in humans and many times are antimicrobial resistant. Production of biofilms further makes matters worse in UTIs. Alternative therapy using bacteriophages was known in the past. This study was aimed to isolate lytic bacteriophages from sewage samples and assess their activity against biofilm of uropathogenic E. coli (UPEC). Lytic phage was isolated from sewage water collected in Jimma town following standard enrichment method against UPEC. E. coli was isolated from UTI suspect patients using standard protocol. Microtiter plate technique was used to determine bacterial biofilm formation. Biofilm degrading efficacy of phage was assessed by treating biofilm developed on cover slip with standardized number of lytic phages or gentamicin compared with the control E. coli (untreated cells). Of 30 UPEC strains isolated from patients, 29 (96.6%) of them displayed biofilm forming phenotype. The strains with strongly biofilm positive were 76.6%. Generally, antibiotic resistance for biofilm producing E. coli was found to be high. Virulent phage (FJS4) was isolated which was effective against a strong biofilm former UPEC strain. Application of FJS4 phage or gentamicin to established biofilms have caused significant reduction of the cells within 3 hours of application and almost complete eradication of the cells within 36 hrs of incubation at 37°C. These results uphold the efficacy of phage against biofilm of UPEC and suggest that FJS4 phage may be a potential therapeutic alternative to antimicrobials on inanimate and animate surfaces.


Keywords


Bacteriophage; Biofilm; Lytic Phage; Uropathogenic E. coli

References


Anderson G, Martin SM, Hultgren SJ. Host subversion by formation of intercellular bacterial communities in the urinary tract. Microbes Infection. 2004, 6:1094-1101

Rodriguez-Bano J, Navarro MD, Romero L, Romero L, Muniain MA, Perea EJ. Clinical and molecular epidemiology of extended-spectrum beta-lactamase-producing Escherichia coli as a cause of nosocomial infection or colonization: implications for control. Clin Infect Dis. 2006, 42: 37-45

Foxman B. Epidemiology of urinary tract infections: Incidence, morbidity, and economic costs. Dis-a-Mon. 2003, 49:53-70

Bower JM, Danelle SE, Matthew AM. Covert operations of uropathogenic Escherichia coli within the urinary tract. Traffic. 2005, 6:18-31

Mulvey MA, Joel DS, Juan JM, Scott JH. Bad bugs and beleaguered bladders: interplay between uropathogenic Escherichia coli and innate host defenses. Proc Natl Acad Sci USA. 2000, 97: 8829-8835

Anderson GG, Palermo JJ, Schilling JD, Roth R, Heuser J, Hultgren SJ. Intracellular bacterial biofilm like pods in urinary tract infections. Science. 2003, 301:105-107

Spoering AM, Lewis K. Biofilm and planktonic cells of Pseudomonas aeruginosa have similar resistance to killing by antimicrobials. J Bacteriol. 2001, 138:6746-6751

Costerton JW, Cheng KJ, Geesey GG. Bacterial biofilms in nature and disease. Ann Rev Microbiol. 1987, 41:435-464

Crouzet M, Claverol S, Lomenech AM, et al. Pseudomonas aeruginosa cells attached to a surface display a typical proteome early as 20 minutes of incubation. PLoS One. 2017, 12:e0180341

Coelho J, Woodford N, Turton J, Livermore DM. Multiresistant Acinetobacter in the UK: how big a threat? J Hosp Infect. 2004, 58: 167-9

Spellberg B, Powers JH, Brass EP, Miller LG, Edwards Jr JE. Trends in antimicrobial drug development: implications for the future. Clin Infect Dis. 2004, 38:1279-86

Czaja CA, Hooton TM. Update on acute uncomplicated urinary tract infection in women. Postgrad Med. 2006, 119: 39-45

Kutter E, Sulakvelidze A. Bacteriophage: Biology and Applications. CRC Press, Inc, Boca Raton, Fla. p. 437-494; 2005

Lacroix-Gueu P, Briandet R, Leveque-Fort S, Bellon-Fontaine MN, Fontaine-Aupart MP. In situ measurements of viral particles diffusion inside mucoid biofilms. C R Biol. 2005, 328: 1065-1072

Hanlon GW, Denyer SP, Ollif CJ, Ibrahim LJ. Reduction in exo-polysaccharide viscosity as an aid to bacteriophage penetration through Pseudomonas aeruginosa biofilms. Appl Environ Microbiol. 2001, 67: 2746-2753

Sillankorva S, Oliveira R, Vieira MJ, Sutherland IW, Azeredo J. Bacteriophage φS1 infection of Pseudomonas fluorescens planktonic cells versus biofilms. Biofouling. 2004, 20:133-138

Curtin JJ, Donlan RM. Using bacteriophages to reduce formation of catheter associated biofilms by Staphylococcus epider¬midis. Antimicrob Agents Chemother. 2006, 50: 1268-1275

Sharp R. Bacteriophages: biology and history. J Chem Tech¬nol Biotechnol. 2001, 76: 667-672.

Ashelford KE, Day MJ, Fry JC. Elevated abundance of bacteriophage infecting bacteria in soil. Appl Environ Microbiol. 2003, 69: 285-289

Leta A, Yohannes M, Kassa T. Assessment of Therapeutic Potential of Bacteriophages to Control Escherichia Coli Infection in Swiss Mice Model. Ethiop J Sci & Technol. 2017, 8: 73-83

Jimma City Administration Strategic Plan (JCASP). Oromia National Regional State, Jimma City Administration Strategic Plan, Jimma, Ethiopia; 2006

ECSA. Ethiopian Central Statistical Agency. Population and Housing Census of Ethiopia administrative report: The Projected result for Oromia Region. Addis Ababa, 2010

Cheesbrough M. District laboratory practice in tropical countries, part II. 2nd edition: England: Butterworth-Heineman LTD, pp105-15; 2006

Collee JG, Fraser AG, Marmion BP, Simmons A. (eds). Mackie and Mc Cartney practical medical microbiology. 14th Ed. New York: Churchill Livingstone; 1996

Hughes KA, Sutherland IW, Clark J, Jones MV. Biofilm susceptibility to bacteriophage attack: the role of phage-borne polysaccharide depolymerase. Microbiology.1998, 144: 3039-3047

Stepanović S, Vuković D, Hola V, Di Bonaventura G, Djukić S, Ćirković I. Quantification of biofilm in microtiter plates: Overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci. Apmis. 2007, 115:891-9

Bauer AW Kirby WMM Sherris JC Turck M. Antibiotic susceptibility testing by standard single disc method. A J Clin Pathol. 1969, 45: 493-496

CLSI. Performance Standards for Antimicrobial Susceptibility Testing. Clinical and Laboratory Standards Institute, Twenty-seventh Information Supplement, 2016; 37(1), M100-S26:1-249

Twest R, Kropinski AM. Bacteriophage enrichment from water and soil: Methods in molecular biology 501. 1:15-32. In: Clokie MRJ, Kropinski AM, editors. Bacteriophages methods and protocols: isolation, characterization, and interactions. New York Springer protocols: Humana Press; 2009

Cerveny KE, DePaola A, Duckworth DH, Gulig PA. Phage therapy of local and systemic disease caused by Vibrio vulnificus in iron-dextran-treated mice. Infect Immun. 2002, 70:6251-6262

Kumari S, Harjai K, Chhibber S. Isolation and characterization of Klebsiella pneumoniae specific bacteriophages from sewage samples. Folia Microbiol. 2010, 55:221-227

Schuch R, Nelson D, Fischetti VA. A bacteriolytic agent that detects and kills Bacillus anthracis. Nature. 2002, 418:884-889

Adams, M.H. Methods of study of bacterial viruses, In: Bacteriophages. London, United Kingdom: Interscience Publishers;1959

Verma V, Harjai K, Chhibber S. Characterization of a T7–like lytic bacteriophage of Klebsiella pneumoniae B5055: A potential therapeutic agent. Curr Microbiol. 2009, 59: 274-281

Kibret M, Abera B. Prevalence and antibiogram of bacterial isolates from urinary tract infections at Dessie Health Research Laboratory, Ethiopia. Asian Pac J Trop Biomed. 2014, 4:164-8

Rijavec M, Müller-Premru M, Zakotnik B, Zgur-Bertok D. Virulence factors and biofilm production among Escherichia coli strains causing bacteraemia of urinary tract origin. J Med Microbiol. 2008, 57: 1329-1334

Ackermann HW. Classification of bacteriophages. The Bacteriophages, Ed. Calendar R, ISBN 0-19-514850-9, New York, USA, Oxford University Press; 2006

Jamalludeen N, Kropinski AM, Johnson RP, Lingohr E, Harel J, Gyles CL. Complete Genomic Sequence of Bacteriophage φEcoM-GJ1, a Novel Phage That Has Myovirus Morphology and a Podovirus-Like RNA Polymerase. Appl Environ Microbiol. 2008, 74(2): 516-525

Merril R, Scholl D, Adhya L. The prospect for Bacteriophage therapy in Western medicine. Nat Rev Drug Discov. 2003, 2:489-497

Sulakvelidze A, Alavidze Z, Morris JG Jr. Bacteriophage therapy. Antimicrob Agents Chemother. 2001, 45:649-659


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