Document Type : Original Article

Authors

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

2 Department of Pathology, Faculty of Veterinary Sciences, University of Veterinary and Animal Sciences, Lahore, Pakistan

3 National Reference Laboratory for Poultry Diseases, Animal Sciences Institute, National Agricultural Research Council, Islamabad, Pakistan

Abstract

The emergence and spread of multidrug resistance among pathogens of the agro-food sector is increasing at an alarming rate, which has directed attention to the search for alternative to antibiotic therapy. The present work studied the physiological and population dynamics of lytic bacteriophages against avian-adapted Salmonella. Out of 28 positive samples, four bacteriophage isolates (SalØ-ABF37, SalØ-RCMPF12, SalØ-MCOH26, SalØ-DNLS42) were selected based on their ability to clearly lyse bacterial test strains. The isolates propagated were active against closely related D1 serotypes, i.e., S. Enteritidis and S. Typhimurium, with no heterologous activity against Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 23235. Each of the monophage suspension and cocktail efficiently suppressed the bacterial count from exponential increase in comparison to the untreated bacterial control. The bacterial turbidity was recorded as 0.244 at λ600 during 400 min of co-incubation, in contrast to bacterial control showing λ600 = 0.669. The latent period was recorded to be 25, 35, 25 and 30 for SalØ-ABF37, SalØ-RCMPF12, SalØ-MCOH26 and SalØ-DNLS42, with 73.00, 97.00, 132 and 75.00 PFU cell-1, respectively. The highest lytic activity was seen at 37.00 ˚C - 42.00 ˚C, with phage particle count being fairly stable at pH 3.00 - 9.00. Each of the isolates possessed dsDNA by being resistant to RNase A. The current study concludes that lytic phages are promising alternative to combat multidrug resistant superbugs. The physiological characterization and bacterial growth inhibition are important parameters in standardization of phage therapy.

Keywords

  1. Organization of economic co-operation and development website. OECD–FAO agricultural outlook. Meat. (6). 2020–2029. Available at https://www.oecd-ilibrary.org/agriculture-and-food/oecd-fao-agricultural -outlook-20202029_1112c23b- Accessed September 11, 2022.
  2. Petracci M, Soglia F, Madruga M, et al. Wooden‐breast, white striping, and spaghetti meat: causes, consequences and consumer perception of emerging broiler meat abnormalities. Compr Rev Food Sci Food Saf 2019; 18(2): 565-583.
  3. Mohsin M, Van Boeckel TP, Saleemi MK, et al. Excessive use of medically important antimicrobials in food animals in Pakistan: a five-year surveillance survey. Glob Health Action. 2019; 12(sup1):1697541. doi: 10.1080/16549716.2019.1697541.
  4. Żbikowska K, Michalczuk M, Dolka B. The use of bacteriophages in the poultry industry. Animals (Basel) 2020; 10(5): 872. doi: 10.3390/ani10050872.
  5. Castro-Vargas RE, Herrera-Sánchez MP, Rodríguez-Hernández R, et al. Antibiotic resistance in Salmonella isolated from poultry: A global overview. Vet World 2020;13(10): 2070-2084.
  6. O’Neill, J. Antimicrobial resistance: Tackling a crisis for the health and wealth of nations. Review on antimicrobial resistance. Available at https://amr-review.org/sites/default/files/AMR Review Paper - Tackling a crisis for the health and wealth of nations_1.pdf. Accessed October 11, 2022.
  7. Fernández L, Gutiérrez D, Rodríguez A, et al. Application of bacteriophages in the agro-food sector: a long way toward approval. Front. Cell Infect Microbiol 2018; 8: 296. doi: 10.3389/fcimb.2018.00296.
  8. Rahaman MT, Rahman MM, Rahman MB, et al. Poultry Salmonella specific bacteriophage isolation and characterization. Bangl J Vet Med 2014; 12(2): 107-114.
  9. Bao H, Zhang P, Zhang H, et al. Bio-control of Salmonella Enteritidis in foods using bacteriophages. Viruses 2015; 7(8): 4836-4853.
  10. Shende RK, Hirpurkar SD, Sannat C, et al. Isolation and characterization of bacteriophages with lytic activity against common bacterial pathogens. Vet World 2017; 10(8): 973-978.
  11. Kwon HJ, Cho SH, Kim TE, et al. Characterization of a T7-like lytic bacteriophage (φSG-JL2) of Salmonella enterica serovar Gallinarum biovar Gallinarum. Appl Environ Microbiol 2008; 74(22): 6970-6979.
  12. Hamza A, Perveen S, Abbas Z, et al. The lytic SA phage demonstrate bactericidal activity against mastitis causing Staphylococcus aureus. Open Life Sci 2016; 11(1): 39-45.
  13. Toro H, Price SB, McKee AS, et al. Use of bacteriophages in combination with competitive exclusion to reduce Salmonella from infected chickens. Avian Dis 2005; 49(1): 118-124.
  14. 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.
  15. Sadekuzzaman M, Mizan MFR, Yang S, et al. Application of bacteriophages for the inactivation of Salmonella in biofilms. Food Sci Technol Int 2018; 24(5): 424-433.
  16. Zhang J, Hong Y, Fealey M, et al. Physiological and molecular characterization of Salmonella bacterio-phages previously used in phage therapy. J Food Prot 2015; 78(12): 2143-2149.
  17. Parra, B, Robeson J. Selection of polyvalent bacteriophages infecting Salmonella enterica serovar Cholerasuis. Electron J Biotechnol 2016;21: 72-76.
  18. Agyare C, Boamah VE, Zumbi CN, et al. Antibiotic use in poultry production and its effects on bacterial resistance. In: Kumar Y (Ed). Antimicrobial resistance - A Global threat. London, UK: Intechopen 1-19.
  19. Medeiros MA, de Oliveira DC, Rodrigues DP, et al. Prevalence and antimicrobial resistance of Salmonella in chicken carcasses at retail in 15 Brazilian cities. Rev Panam Salud Publica 2011; 30(60): 555-560.
  20. Ackermann HW. Bacteriophage taxonomy. Microbiol Aust 2011; 32(2): 90-94.
  21. Bielke LR, Higgins SE, Donoghue AM, et al. Use of wide-host-range bacteriophages to reduce Salmonella on poultry products. Int J Poult Sci 2007; 6(10): 754-757.
  22. Borie C, Albala I, Sánchez P, et al. Bacteriophage treatment reduces Salmonella colonization of infected chickens. Avian Dis 2008; 52(1): 64-67.
  23. Nale JY, Vinner GK, Lopez VC, et al. An optimized bacteriophage cocktail can effectively control Salmonella in vitro and in Galleria mellonella. Front. Microbiol 2021; 11:609955. doi: 10.3389/fmicb. 2020.609955.
  24. Islam MS, Hu Y, Mizan MFR, et al. Characterization of Salmonella Phage LPST153 That Effectively Targets Most Prevalent Salmonella Microorganisms 2020; 8(7): 1089. doi: 10.3390/microorganisms8071089.
  25. Hong SS, Jeong J, Lee J, et al. Therapeutic effects of bacteriophages against Salmonella Gallinarum infection in chickens. J Microbiol Biotechnol 2013; 23(10): 1478-1483.
  26. Hungaro HM, Santos Mendonça RC, 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.
  27. Ngangbam AK, Devi NB. Molecular characterization of Salmonella bacteriophages isolated from natural environment and its potential role in phage therapy. Bangla J Microbiol 2012; 29(1): 33-36.
  28. Fiorentin L, Vieira ND, Barioni Júnior W, et al. In vitro characterization and in vivo properties of Salmonella lytic bacteriophages isolated from free-range layers. Braz J Poult Sci 2004; 6(2): 121-128.