Novel Phage Cocktail for the Treatment of Bacteria Causing Chronic Suppurative Otitis Media doi.org/10.26538/tjnpr/v4i10.4
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Abstract
Chronic suppurative otitis media (CSOM) is a chronic inflammation of the middle ear cavity and mastoid cavity. It occurs in any community in a percentage of 0.5-33% for all age group. The objective of this study was to isolate and prepare phage cocktail active against bacteria causing chronic suppurative otitis media from sewage water. Bacteria were isolated from non-drug responsive cases of CSOM. A total of 73 ear swabs were collected from patients of CSOM and showed bacterial growth in 63 (86.3%) and fungal growth in 10 (13.7%). The most prevalent bacterium was Pseudomonas aeruginosa which was found in 33 swabs (45.2%), followed by Staphylococcus aureus in 15 (20.5%). Species of Klebsiella pneumoniae, Escherichia coli, Proteus mirabilis, and Moraxella catarrhalis were isolated in 5 (6.8%), 5 (6.8%), 4 (5.5%) and 1 (1.3%), respectively. The most potent phages were 7, 3, 1, 2, 1 and 1 for species of P.
aeruginosa, S. aureus, K. pneumoniae, E. coli, P. mirabilis and M. catarrhalis, respectively. Phages were isolated after enrichment of each CSOM bacterial pathogen with sewage water by spot lysis method and characterized morphologically by top agarose plaque assay. Most virulent isolated phages were selected for formulation of phage cocktail against isolated bacterial pathogens of CSOM. A number of 17 phages cocktail were formulated and showed 60-100% inhibition on all tested isolates of P. aeruginosa, S. aureus, K. pneumoniae, E. coli, P. mirabilis and M. catarrhalis in a growth inhibition assay.
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References
World Health Organization. Chronic suppurative otitis media : burden of illness and management options. Geneve :World Health Organization. 2004.
Abraham ZS, Ntunaguzi D, Kahinga AA, Mapondella KB, Massawe ER, Nkuwi EJ, NKya A. Prevalence and etiological agents for chronic suppurative otitis media in a tertiary hospital in Tanzania. BMC Res Notes. 2019; 12:429.
Kenna MA. Treatment of chronic suppurative otitis media. Otolaryngol Clin North Am. 1994; 27(3):457-472.
Dubey SP and Larawin V. Complications of Chronic Suppurative Otitis Media and Their Management. The Laryngoscope 2007; 117:264-267.
Adhikari P, Sinha BK, Pokhrel NR, Kharel B, Aryal R, et al. Prevalence of chronic suppurative otitis media in school children of kathmandu district. J Inst Med. 2007; 29(3):10-12.
John NM, Balachandrakurup N, Varghese S. A prospective study on microbiological profile and antimicrobial resistance pattern in chronic suppurative otitis media at a rural centre. Int J Otorhinolaryngol Head Neck Surg. 2020; 6(3):542-546.
Lee SK, Lee MS, Jung SY, Byun JY, Park MS, Yeo SG. Antimicrobial resistance of Pseudomonas aeruginosa from otorrhea of chronic suppurative otitis media patients. Otolaryngol Head Neck Surg. 2010; 143(4):500-505.
Michael RL, Pawlowski KS, Luong A, Furze AD, Roland PS. Biofilm presence in humans with chronic suppurative otitis media. Otorhinolaryngol Head Neck Surg. 2009; 141(5):567-571.
Abedon ST. Ecology of Anti-Biofilm Agents II: Bacteriophage Exploitation and Biocontrol of Biofilm Bacteria. Pharm (Basel). 2015; 8(3):559–589.
Gutiérrez D, Rodríguez-Rubio L, Martínez B, Rodríguez A, García P. Bacteriophages as Weapons Against Bacterial Biofilms in the Food Industry. Front Microbiol. 2016; 7:825.
Summers WC. Bacteriophage Therapy. Ann Rev Microbiol. 2001; 55(1):437-451.
Duplessis C, Biswas B, Hanisch B, Perkins M, Henry M, Quinones J, Wolfe D, Estrella L, Hamilton, T . Refractory pseudomonasbacteremia in a 2-year-old sterilized by bacteriophage therapy. J Pediatric Infect Dis Soc. 2017; 7:253–256.
Khawaldeh A, Morales S, Dillon B, Alavidze Z, Ginn AN, Thomas L,Chapman, SJ, Dublanchet A, Smithyman A, Iredell JR. Bacteriophage therapy for refractory Pseudomonas aeruginosa urinary tract infec-tion. J Med Microbiol. 2011; 60:1697–1700.
Sarker SA, Sultana S, Reuteler G, Moine D, Descombes P, Charton F, Bourdin G, McCallin S, Ngom-Bru C, Neville T, Akter M, Huq S, Qadri F, Talukdar K, Kassam M, Delley M, Loiseau C, Deng Y, El Aidy S, Berger B, Brüssow H. Oral Phage Therapy of Acute Bacterial Diarrhea With Two Coliphage Preparations: A Randomized Trial in Children From Bangladesh. E BioMed. 2016; 4:124‐137.
Petrovic Fabijan A, Lin RCY, Ho J, Maddocks S, Zakour NLB, Iredell JR. Safety of bacteriophage therapy in severe Staphylococcus aureus infection. Nat Microbiol. 2020; 5:465–472.
Jault P, Leclerc T, Jennes S, Pirnay JP, Que YA, Resch G, Rousseau AF, Ravat F, Carsin H, Le Floch R, Schaal JV, Soler C, Fevre C, Arnaud I, Bretaudeau L, Gabard J. Efficacy and tolerability of a cocktail of bacteriophages to treat burn wounds infected by Pseudomonas aeruginosa
(PhagoBurn): a randomised, controlled, double-blind phase 1/2 trial. Lancet Infect Dis. 2018; 19:35–45.
Clinical and Laboratory Standards Institute (CLSI). Performance standards for antimicrobial susceptibility testing. 20th informational supplement. M 100-S20, Wayne, Pennsylvania; 2014; 30 (1).
Kumari S, Harjai K, Chhibber S. Isolation and characterization of Klebsiella pneumoniae specific bacteriophages from sewage samples. Folia Microbiol. 2010; 55:221–227.
Tan GH, Nordin MS, Napsiah AB. Isolation and characterization of lytic bacteriophages from sewage water (Pengasingan dan pencirian bakteriofaj daripada air kumbahan). J Trop Agric Food Sci. 2008; 36(2):1-5.
Jassim SAA, Abdulamir AS, Abu Bakar F. Novel phagebased bio-processing of pathogenic Escherichia coli and its biofilms. World J Microbiol Biotechnol. 2012; 28:47–60.
Knezevic P and Petrovic O. A colorimetric microtiter plate method for assessment of phage effect on Pseudomonas aeruginosa biofilm. J Microbiol Meth. 2008; 74(2-3):114- 118.
Abdelshafy IA, Haleem AA, Khalil YA, Ghazal AA, Gaballah A, Saied A. Microbiology of Chronic Suppurative Otitis Media, Study of the Role of Bacterial Biofilm and Fungal Infection. J Otolaryngol ENT Res. 2015; 3(1):00051.
ittal R, Lisi CV, Gerring R, Mittal J, Mathee K, Narasimhan G, Azad RK, Yao Q, Grati M, Yan D, Eshraghi AA, Angeli SI, Telischi FF, Liu XZ. Current concepts in the pathogenesis and treatment of chronic suppurative otitis media. J Med Microbiol. 2015; 64(10):1103–1116.
DeAntonio R, Yarzabal JP, Cruz JP, Schmidt JE, Kleijnen J. Epidemiology of otitis media in children from developing countries: A systematic review. Int J Ped Otorhinolaryngol. 2016; 85:65–74.
Addas F, Algethami M, Mahmalji N, Zakai S, Alkhatib T. Bacterial Etiology and Antimicrobial Sensitivity Patterns of Ear Infections at King Abdulaziz University Hospital, Jeddah, Saudi Arabia. J Nat Sci Med. 2019; 2:147-152.
Pang Z, Raudonis R, Glick BR, Lin T, Cheng Z. Antibiotic resistance in Pseudomonas aeruginosa: mechanisms and alternative therapeutic strategies. Biotechnol Adv. 2019; 37(1):177-192.
Pires DP, Melo LDR, Vilas Boas D, Sillankorva S, Azeredo J. Phage therapy as an alternative or complementary strategy to prevent and control biofilm-related infections. Curr Opin Microbiol. 2017; 39:48- 56.
Rakhuba DV, Kolomiets EI, Szwajcer Dey E, Novik G.I. Bacteriophage Receptors, Mechanisms of Phage Adsorption and Penetration into Host Cell. Pol J Microbiol. 2010; 59(3):145-155.
Randall-Hazelbauer L and Schwartz M. Isolation of the bacteriophage lambda receptor from Escherichia coli. J Bacteriol. 1973; 116:1436-1446.
Yu F and Mizushima S. Roles of lipopolysaccharide and outer membrane protein OmpC of Escherichia coli K12 in the receptor function for bacteriophage T4. J Bacteriol. 1982; 51:423-434.
Hashemolhosseini S, Holmes Z, Mutschler B, Henning U. Alterations of receptor specificities of coliphages of the T2 family. J Mol Biol. 1994; 240:105-110.
Kaabi SAG and Musafer HK. An experimental mouse model for phage therapy of bacterial pathogens causing bacteremia. Microb Pathog. 2019; 137:103770.
Kaabi SAG and Musafer HK. New Phage cocktail against infantile Sepsis bacteria. Microb Pathog. 2020; 148:104447.