Abdallah M. A. Merwad, Mahmoud E. F. Abdel-Haliem


This study targeted isolation and characterization of phage against multidrug resistant (MDR) Salmonella Typhimurium and Salmonella Enteritidis recovered from broilers and to evaluate the lytic effect of the phage on growth of Salmonella serovars. Salmonella isolates were recovered from caecal contents, liver and breast meat of broiler chickens from retail outlets at Sharkia Governorate, Egypt. Salmonella Typhimurium (n=14) and S. Enteritidis (n=11) were tested for their antimicrobial susceptibilities against 15 antimicrobials by disc diffusion method. Isolates of S. Typhimutium and S. Enteritidis were 100% resistant to seven antimicrobial agents. The phage was isolated from Zagazig sewage water by spot test and double over layer agar assay. The phage designated as phiSalmchick1showed an icosahedral head and contractile tail structure in electron microscopy, indicating a member of the family Myoviridae. The phage was a polyvalent infecting a wide host range of all MDR strains of S. Typhimurium, S. Enteritidis, S. Paratyphi, E. coli serotypes O26 and O168 and Klebsiella pneumoniae. Myovirus phage had burst size of 100 plague forming unit (PFU)/cell with latent period of 60 min. The phage genome had double-stranded DNA by molecular analysis. The lytic effect of phiSalmchick1 phage was in vitro assessed on growth of S. Typhimurium and S. Enteritidis isolates by measuring the optical density (OD) of the liquid media during Salmonella growth at 37 °C and the multiplicity of infection (MOI) was equal to 1.0. Significant reductions were observed in OD of S. Typhimurium and S. Enteritidis treated with the phage after 24 hrs incubation compared to the controls (P<0.05). The myovirus has a high potential for phage application to control zoonotic and MDR Salmonella serovars isolated from broiler chickens in Egypt.

Key words: multidrug resistant; Salmonella serovars; Myoviridae phage; broiler chickens; in vitro control

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(1) Nagappa K, Tamuly S, Brajmadhuri, Saxena MK, Singh SP. Isolation of Salmonella Typhimurium from poultry eggs and meat of Tarai region of Uttaranchal. Ind J Biotechnol 2007; 6: 407–9.

(2) Bhunia AK. Salmonella enterica. In: Foodborne microbial pathogens: mechanisms and pathogenesis. USA: Springer Science+Business Media, LLC 2008; 201–15.

(3) Naurin S, Islam MA, Khatun MM. Prevalence of Salmonella in apparently healthy chickens in Mymensingh, Bangladesh. Microbes and Health 2012; 1: 30–3.

(4) Saad SM, Nada S, Abd El Sattar SS. Incidence of Salmonella species in chicken cut-up carcasses and chicken products. Benha Vet Med J 2015; 29: 29–35.

(5) Abd El‑Ghany W, El‑Shafii S, Hatem M. A survey on Salmonella species isolated from chicken flocks in Egypt. Asian J Anim Vet Adv 2012; 7: 489–501.

(6) Okeke IN, Laxminarayan R, Bhutta ZA, Duse AG, Jenkins P, O’Brien TF, Pablos-Mendez A, Klugman KP. Antimicrobial resistance in developing countries. Part I: recent trends and current status. Lancet Infect Dis 2005; 5: 481–93.

(7) Mahmud MS, Bari ML, Hossain MA. Prevalence of Salmonella serovars and antimicrobial resistance profiles in poultry of Savar area, Bangladesh. Foodborne Pathog Dis 2011; 8: 1111–8.

(8) Mahony J, McAuliffe O, Ross RP, van Sinderen D. Bacteriophages as biocontrol agents of food pathogens. Curr Opin Biotechnol 2011; 22: 157–63.

(9) Parsley LC, Consuegra EJ, Thomas SJ., Bhavsar J, Land AM, Bhuiyan NN Mazher MA, Waters RJ, Wommack KE, Harper WF Jr, Liles MR.. Census of the viral metagenome within an activated sludge microbial assemblage. Appl Environ Microbiol 2010; 76: 2673–77.

(10) McCallin S, Alam Sarker S, Barretto C, Sultana S, Berger B, Huq S, Krause L, Bibiloni R, Schmitt B, Reuteler G, Brüssow H. Safety analysis of a Russian phage cocktail: from MetaGenomic analysis to oral application in healthy human subjects. Virology 2013; 443: 187–96.

(11) Mahmoud M, Askora A, Barakat AB, Rabie OE, Hassan SE. Isolation and characterization of polyvalent bacteriophages infecting multi drug resistant Salmonella serovars isolated from broilers in Egypt. Int J Food Microbiol 2018; 266: 8–13.

(12) ISO-6579. International organization for standardization: Microbiology of food and animal feeding stuffs-horizontal method for detection of Salmonella spp. 2002.

(13) Andrews WH, Jacobson A, Hammack TS. Salmonella. In: Bacteriological Analytical Manual 1998, 8th ed., Ch. 4. Revision A, Accessed on16-04-2015.

(14) Kauffmann F, Das-Kauffmann W. Antigenic formulas of the Salmonella serovars 8th Edition, 2001, WHO co-operating centre for reference and research on Salmonella.

(15) Bauer AW, Kirby WM, Sherris JC, Turck M. Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol 1966; 45: 493–6.

(16) McFarland J. Nephelometer: an instrument for estimating the number of bacteria in suspensions used for calculating the opsonic index and for vaccines. J Am Med Assoc 1907; 14: 1176–78.

(17) CLSL. Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing; six-teenth informational supplement. M100-S16, Vol. 26, No. 3, Clinical and Laboratory Standards Institute 2016 Wayne, Pennsylvania,19087–1898, USA.

(18) Adams MH. 1959. Bacteriophages. Interscience Publishers, New York.

(19) Carvalho C, Susano M, Fernandes E, Santos S, Gannon B, Nicolau A, Gibbs P, Teixeira P, Azeredo J. Method for bacteriophage isolation against target Campylobacter strains. Lett Appl Microbiol 2010; 50: 192–7.

(20) Abdel-Haliem MEF, Askora A. Isolation and characterization of bacteriophages of Helicobacter pylori isolated from Egypt. Future Virology 2013; 8: 821–6.

(21) Pajunen M, Kiljunen S, Skurnik M. (2000. Bacteriophage phi YeO3-12, specific for Yersinia enterocolitica serotype O: 3, is related to coliphages T3 and T7. J Bacteriol 2000; 182: 5114–20.

(22) Sambrook J, Russell D. Molecular cloning: a laboratory manual, 3rd edn, 2001 Cold Spring Harbor Laboratory Press, Cold Spring Harbor: 44–6.

(23) Yoon SS, Barrangou-Poueys R., Breidt Jr F, Fleming HP. Detection and characterization of a lytic Pediococcus bacteriophage from the fermenting cucumber brine.J. Microbiol. Biotechnol 2007; 17: 262–70.

(24) Bielke LR., Higgins AM., Donoghue AM., Donoghue DJ, Hargis BM. Salmonella host range of bacteriophages that infect multiple genera. Poult Sci 2007; 86: 2536–40.

(25) Parra, B., Robeson, J., 2016. Selection of polyvalent bacteriophages infecting Salmonella enterica serovar Choleraesuis. Electron J Biotechnol 2016; 21: 72–6.

(26) Duc HM, Son HM, Honjoh K, Miyamoto T. Isolation and application of bacteriophages to reduce Salmonella contamination in raw chicken meat. LWT - Food Sci Technol 2018; 91: 353–60.

(27) Parisien, A., Allain B., Zhang J., Mandeville R., Lan CQ, 2008. Novel alternatives to antibiotics: bacteriophages, bacterial cell wall hydrolases, and antimicrobial peptides. J Appl Microbiol 2008; 104: 1–13.

(28) Monteville MR, Ardestani B, Geller BL 1994. Lactococcal bacteriophages require a host cell wall carbohydrate and a plasma membrane protein for adsorption and ejection of DNA. Appl Environ Microbiol 1994; 60: 3204–11.

(29) Kuhn J1, Suissa M, Chiswell D, Azriel A, Berman B, Shahar D, Reznick S, Sharf R, Wyse J, Bar-On T, Cohen I, Giles R, Weiser I, Lubinsky-Mink S, Ulitzur S. A bacteriophage reagent for Salmonella: Molecular studies on Felix 01. Int J Food Microbiol 2002; 74: 217–27.

(30) Hadas H, Einav M, Fishov I, Zaritsky A. 1997. Bacteriophage T4 development depends on the physiology of its host Escherichia coli. Microbiology 1997; 143: 179–85.

(31) Muller-Merbach M, Kohler K, Hinrichs J. Environmental factors for phage induced fermentation problems: replication and adsorption of the Lactococcus lactis phage P008 as influenced by temperature and pH. Food Microbiol 2007; 24: 695–702.

(32) Gill JJ, Hyman P. Phage choice, isolation, and preparation for phage therapy. Curr Pharm Biotechnol 2010; 11: 2–14.

(33) Hudson JA, Billington C, Carey-Smith G, Greening G. 2005. Bacteriophages as biocontrol agents in food. J. Food Prot 2005; 68: 426–37.

(34) Nikkhahi F, Soltan Dallal MM, Alimohammadi M, Rahimi Foroushani A, Rajabi Z, Fardsanei F, Imeni SM, Torabi Bonab P. 2017. Phage therapy: assessment of the efficacy of a bacteriophage isolated in the treatment of salmonellosis induced by Salmonella enteritidis in mice. Gastroenterol Hepatol Bed Bench 2017; 10: 131–6.

(35) Atterbury RJ, Van Bergen MA, Ortiz F, Lovell MA, Harris JA, De Boer A, Wagenaar JA, Allen VM, Barrow PA. Bacteriophage therapy to reduce Salmonella colonization of broiler chickens. Appl Environ Microbiol 2007; 73: 4543–9.



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