Veterinary Research Forum
Vet Res Forum

In-vitro evaluation and efficacy of bacteriophage isolated from commercial poultry farms against Salmonella typhimurium

Document Type : Original Article

Authors

Amina Zaib 1
Ali Ahmad Sheikh 1
Faisal Ayub 1
Muhammad Arshad Durrani 1
Mobashra Mustafa 1
Afifa Shahzad 1
Urooj Ejaz 1
Muhammad Zubair Latif 2
Chamman Zahra 1
Izza Izza 1
Fareeha Zaib 3
Husnain Ahmad 1
Muhammad Jawad Hafeez 1
Muhammad Mujahid 1
Hafiz Muhammad Moavia Atique 1

1 Institute of Microbiology, Faculty of Veterinary Sciences, University of Veterinary and Animal Sciences, Lahore, Pakistan

2 Veterinary Research Institute, Lahore, Pakistan

3 College of Earth and Environmental Sciences, University of the Punjab, Lahore, Pakistan

https://doi.org/10.30466/vrf.2024.2030436.4305
Abstract
Avian Salmonellosis impacts the economy and public health, with chicken products being a major cause of gastroenteritis. Hygiene, immunization and medicines are all used as control techniques. Bacteriophages provide a safe, targeted alternative. In the present study in vitro evaluation of bacteriophages were done against Salmonella typhimurium. Lytic effect of bacteriophages isolated from poultry sludge was checked on culture of S. typhimurium. Stability study was checked at range of temperature and pH. The phages were stable at temperature (30.00 - 50.00 ˚C) and pH (5.00 - 9.00) where best activity was seen at 37.00 ˚C and pH 7.00. In vitro lytic activity was done at (optical density 600 nm) after exposure to bacterial host at different intervals. Multiplicity of Infection of 1.00 was used to check lytic activity of phages which indicated phages were potent enough to infect bacterial cells within their growth cycle. The percentage of unadsorbed phages was determined by bar chart analysis. The genome of three phages was treated with DNase I where they all were sensitive. Later the nucleic acid of phages was digested by restriction endonucleases (EcoRI and HindIII) both of the enzymes produced various restriction sites with different band. The present study proved that the application of bacteriophages in vitro into bacterial system i.e., S. typhimurium was an attractive method in diminishing infection in commercial poultry thus providing exceptional results that could be used on a large scale.

Keywords

Bacteriophages
Poultry
Restriction endonucleases
Salmonella typhimurium
Salmonellosis

Subjects

  1. McQuiston JR, Herrera-Leon S, Wertheim BC, et al. Molecular phylogeny of the Salmonellae: relationships among Salmonella species and subspecies determined from four housekeeping genes and evidence of lateral gene transfer events. J Bacteriol 2008; 190(21): 7060-7067.
  2. Margos G, Ogden NH, Fingerle V, et al. Bacteria of the genus Borrelia. In: Fereidouni S (Ed). Ecology of wild bird diseases, 1st ed Florida, USA, CRC Press 2024; 271-300.
  3. Huang H, Naushad S. Salmonella: perspectives for low-cost prevention, control and treatment. 1st London, UK: IntechOpen 2024; 1-5.
  4. Pande VV, Devon RL, Sharma P, et al. Study of Salmonella Typhimurium infection in laying hens. Front microbiol 2016; 7: 203. doi: 10.3389/fmicb. 2016.00203.
  5. Perez-Lopez A, Behnsen J, Nuccio SP, et al. Mucosal immunity to pathogenic intestinal bacteria. Nat Rev Immunol 2016; 16(3): 135-148.
  6. Castro-Vargas RE, Herrera-Sánchez MP, Rodríguez-Hernández R, et al. Antibiotic resistance in Salmonella spp. isolated from poultry: a global overview. Vet World 2020; 13(10): 2070-2084.
  7. Zdolec N, Kiš M. Meat safety from farm to slaughter—Risk-based control of Yersinia enterocolitica and Toxoplasma gondii. Processes 2021; 9(5): 815. doi: 10.3390/pr9050815.
  8. Perumalla AS, Wythe LA, Ricke S. Probiotics in poultry preharvest food safety: Historical Developments and Current Prospects. In: Callaway TR, Ricke SC (Eds). Direct-fed microbials and prebiotics for animals: Science and mechanisms of action. London, UK: Springer, Cham; 2023:127-166.
  9. Hays S, Page SJ, Buhalis D. Social media as a destination marketing tool: its use by national tourism organizations. Curr Issues Tourism 2012; 16(3): 211-239.
  10. Harada LK, Silva EC, Campos WF, et al. Biotechnological applications of bacteriophages: state of the art. Microbiol Res 2018; 212-213: 38-58.
  11. Kumari P, Chakraborty T, Ghosh S. Advancement of phage therapy approaches in the battle of multi-drug resistance: a review. Int J Life Sci Pharma Res 2023; 13(2): 17-36.
  12. Au A, Lee H, Ye T, et al. Bacteriophages: combating antimicrobial resistance in food-borne bacteria prevalent in agriculture. Microorganisms 2021;10(1): 46. doi: 10.3390/microorganisms10010046.
  13. Farooq T, Hussain MD, Shakeel MT, et al. Deploying viruses against phytobacteria: potential use of phage cocktails as a multifaceted approach to combat resistant bacterial plant pathogens. Viruses 2022; 14(2): 171. doi: 10.3390/v14020171.
  14. Andreatti Filho RL, Higgins JP, Higgins SE, et al. Ability of bacteriophages isolated from different sources to reduce Salmonella enterica serovar enteritidis in vitro and in vivo. Poult Sci 2007; 86(9): 1904-1909.
  15. Atterbury RJ, Van Bergen MA, Ortiz F, et al. Bacteriophage therapy to reduce Salmonella colonization of broiler chickens. Appl Environ Microbiol 2007; 73(14): 4543-4549.
  16. Baran V, Colonna M, Greco V, et al. Reaction dynamics with exotic nuclei. Phys Rep 2005; 410(5-6): 335-466.
  17. Toulon P, Ozier Y, Ankri A, et al. Point-of-care versus central laboratory coagulation testing during haemorrhagic surgery. Thromb Haemost 2009; 101(02): 394-401.
  18. Wong WL, Su X, Li X, et al. Global prevalence of age-related macular degeneration and disease burden projection for 2020 and 2040: a systematic review and meta-analysis. Lancet Glob Health 2014; 2(2): e106-e116.
  19. Gupta K, Hooton TM, Naber KG, et al. International clinical practice guidelines for the treatment of acute uncomplicated cystitis and pyelonephritis in women: a 2010 update by the Infectious Diseases Society of America and the European Society for Microbiology and Infectious Diseases. Clin Infect Dis 2011;52(5): e103-e120.
  20. Hungaro HM, Mendonça RCS, Gouvêa DM, et al. Use of bacteriophages to reduce Salmonella in chicken skin in comparison with chemical agents. Food Res Int 2013; 52(1): 75-81.
  21. Akhtar M, Viazis S, Diez-Gonzalez F. Isolation, identification and characterization of lytic, wide host range bacteriophages from waste effluents against Salmonella enterica serovars. Food control 2014; 38(1): 67-74.
  22. Pereira C, Moreirinha C, Lewicka M, et al. Bacteriophages with potential to inactivate Salmonella Typhimurium: use of single phage suspensions and phage cocktails. Virus Res 2016; 220: 179-192.
  23. Rahaman MT, Rahman M, Rahman MB, et al. Poultry Salmonella specific bacteriophage isolation and characterization. Bangl J Vet Med 2014; 12(2): 107-114
  24. Shaheen AY, Sheikh AA, Rabbani M, et al. Isolation and physicochemical characterization of brucella phages. J Anim Plant Sci 2021; 31(5): 1277-1286
  25. Bao H, Zhang H, Wang R. Isolation and characterization of bacteriophages of Salmonella enterica serovar Pullorum. Poult Sci 2011; 90(10): 2370-2377.
  26. Santos SB, Carvalho C, Azeredo J, et al. Population dynamics of a Salmonella lytic phage and its host: implications of the host bacterial growth rate in modelling. PloS One 2014; 9(7): e102507. doi: 10.1371/journal.pone.0102507.
  27. Salmonellosis in animals website. Salmonella - a re-emerging pathogen. Available at: www.intechopen. com/chapters/58593. Accessed April 30, 2025.
  28. He Y, Wang J, Zhang R, et al. Epidemiology of foodborne diseases caused by Salmonella in Zhejiang Province, China, between 2010 and 2021. Front Public Health 2023; 11:1127925. doi: 10.3389/fpubh.2023.1127925.
  29. Antunes P, Mourão J, Campos J, et al. Salmonellosis: the role of poultry meat. Clin Microbiol Infect 2016; 22(2): 110-121.
  30. McDermott PF, Zhao S, Tate H. Antimicrobial resistance in nontyphoidal Salmonella. Microbiol Spectr 2018; 6(4): 10.1128/microbiolspec.arba-0014-2017. doi: 10.1128/microbiolspec.ARBA-0014-2017.
  31. Mburu SN. Antimicrobial resistance in non-typhoidal Salmonella recovered from selected food animals in Kenya. MSc. Thesis, Jomo Kenyatta University of Agriculture and Technology. Juja, Kenya: 2020.
  32. Bekoz Yarar M, Turkyilmaz MK, Turkyilmaz S. Investigation of integron, antimicrobial resistance and virulence gene profiles of Salmonella enterica subspecies enterica serovar infantis isolates obtained from broiler chickens. Isr J Vet Med 2023; 78(4): 12-27.
  33. Han JE, Kim JH, Hwang SY, et al. Isolation and characterization of a Myoviridae bacteriophage against Staphylococcus aureus isolated from dairy cows with mastitis. Res Vet Sci 2013; 95(2): 758-763.
  34. Chhibber S, Kaur P, Gondil VS. Simple drop cast method for enumeration of bacteriophages. J Virol Methods 2018; 262: 1-5.
  35. Teklemariam AD. Isolation and characterization of bacteriophages for some multi drug-resistant bacteria. Ph.D. Thesis, King Abdulaziz University. Jeddah, Saudi Arabia: 2023.
  36. Singh BR, Sinha D, Agrawal RK, et al. Alternative Approaches to Mitigate Antimicrobial Drug Resistance. Izatnagar, India: ICAR-Indian Veterinary Research Institute (IVRI), 2021: 1-110.
  37. Yang Q, Ding Y, Nie R, et al. Characterization of a novel T7-like Salmonella Typhimurium (ATCC13311) bacteriophage LPST144 and its endolysin. LWT 2020; 123: 109034. doi: 10.1016/j.lwt.2020.109034.
  38. Żbikowska K, Michalczuk M, Dolka B. The use of bacteriophages in the poultry industry. Animals (Basel) 2020; 10(5): 872. doi: 10.3390/ani10050872.
  39. Hyman P, Abedon ST. Practical methods for determining phage growth parameters. Methods Mol Biol 2009; 501: 175-202.
  40. Gautam A. DNA and RNA Isolation Techniques for Non-experts. Cham, Switzerland: Springer 2022; 1-273
  41. Muhilarasi S. Partial genomic characterization and isolation of endolysin from bacteriophage SFN6B against Shigella flexneri: BSc Thesis, Universiti Tunku Abdul Rahman. Petaling Jaya, Malaysia: 2015.
  42. Aaron JA. Isolation and characterization of bacteriophages infecting UTI-associated bacteria and evaluation of phage-derived proteins as potential therapeutic agents and diagnostic probes. MSc. Thesis, Stellenbosch University. Stellenbosch, South Africa: 2023.
  43. Dlamini SB, Gigante AM, Hooton SPT, et al. Efficacy of different encapsulation techniques on the viability and stability of diverse phage under simulated gastric conditions. Microorganisms 2023; 11(10): 2389. doi: 10.3390/microorganisms11102389.
Veterinary Research Forum
Volume 16, Issue 5
May 2025
Pages 267-275

  • Receive Date 01 June 2024
  • Revise Date 05 November 2024
  • Accept Date 06 November 2024

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