Application of bacteriophages to control Salmonella Enteritidis in raw eggs

Document Type : Short paper


1 Nitte (Deemed to be University), Nitte University Centre for Science Education and Research, Deralakatte, Mangaluru, 575 018, India

2 Virus Research and Diagnostic Laboratory (VRDL), Shimoga Institute of Medical Sciences (SIMS), Shivamogga-577 201, India


Background: Salmonella, a bacterial genus of more than 2500 serotypes is considered as the most significant foodborne pathogen causing infections in humans and animals. Increased antimicrobial resistance and persistence of antimicrobial residues in food matrices warrants the need for alternative infection management strategies. Aims: The present study aimed to isolate and evaluate the lytic activity of bacteriophage against Salmonella. Methods: Twenty-eight Salmonella isolates obtained from the poultry sources were screened for antibiotic sensitivity. Poultry slaughterhouse wastewater was used for the isolation of phage. Host range and random amplified polymorphic DNA (RAPD) are vital tools used for differentiating the phage. Results: The isolates showed a high degree of resistance to nalidixic acid (71%), tetracycline (71%), nitrofurantoin (50%), and ampicillin (43%). Five lytic phages are specific for Salmonella spp. were isolated and characterized by RAPD. In the colony forming unit (CFU) reduction assay, the highest activity of phage was observed at 0.01 multiplicity of infection (MOI) within 2 h after the addition of phage. PSE5 at 0.01 MOI was administered to Salmonella Enteritidis seeded on the surface of the chicken egg by immersion method. The results indicated that administration of phage reduced recoverable Salmonella by 2 × 106 CFU/ml relative to the phage-excluded control. Conclusion: The results presented here suggested the application of the bacteriophage treatment has the potential to be used as an alternative strategy to prevent Salmonella infection in poultry farms to prevent vertical transmission of the pathogen.


Main Subjects

Adams, MH (1959). Enumeration of bacteriophage particles. Bacteriophages, London, Interscience Publishers, Ltd., PP: 27-34.
Adriaenssens, EM; vanVaerenbergh, J; Vandenheuvel, D; Dunon, V; Ceyssens, PJ; deProft, M; Kropinski, AM; Noben, JP; Maes, M and Lavigne, R (2012). T4-related bacteriophage LIME stone isolates for the control of soft rot on potato caused by Dickeya solani. PLoS One., 7: e33227.
Andino, A and Hanning, I (2015). Salmonella enterica: survival, colonization, and virulence differences among serovars. Sci. World J., doi: 0.1155/2015/520179.
Ashwale, JK; Rohde, M; Rohde, C; Bunk, B; Sproer, C; Boga, HI; Klenik, HP and Wittmann, J (2019). Isolation, characterization and analysis of bacteriophages from the haloalkaline lake Elmenteita, Kenya. PLOS One. 10: e0004813.
Babickova, J and Gardlik, R (2015). Pathological and therapeutic interactions between bacteriophages, microbes and the host in inflammatory bowel disease. World J. Gastroenterol., 21: 11321-11330.
Besler, I; Sazinas, P; Harrison, C; Gannon, L; Redowell, T; Michniewski, S; Hooton, SP and Milard, A (2020). Genome sequence and characterization of Coliphage vB_Eco_SLUR29. Phage. 1: 38-44.
Briers, Y; Walmagh, M; Van Puyenbroeck, V; Cornelissen, A; Cenens, W; Aertsen, A; Oliveira, H; Azeredo, J; Verween, G; Pirnay, J; Miller, S; Volckaert, G and Lavigne, R (2014). Engineered endolysin-based “Artilysins” to combat multidrug-resistant Gram-negative pathogens. mBio., 5: e01379-14.
Buncic, S and Sofos, J (2012). Interventions to control Salmonella contamination during poultry, cattle and pig slaughter. Food Res. Int., 45: 641-655.
Campos, J; Mourão, J; Peixe, L and Antunes, P (2019). Non-typhoidal Salmonella in the pig production chain: A comprehensive analysis of its impact on human health. Pathogens. 8(1), 19.
Crum-Cianflone, NF (2008). Salmonellosis and the gastrointestinal tract: more than just peanut butter. Curr. Gastroenterol. Rep., 10: 424-431.
El-Dougdoug, NK; Cucic, S; Abdelhamid, AG; Brovko, L; Kropinski, AM; Griffiths, MW and Anany, H (2019) Control of Salmonella Newport on cherry tomato using a cocktail of lytic bacteriophages. Int. J. Food Microbiol., 293: 60-71.
Endersen, L; O’Mahony, J; Hill, C; Ross, RP; McAuliffe, O and Coffey, A (2014). Phage therapy in the food industry. Annu. Rev. Food Sci. Technol., 5: 327-349.
Hooton, SPT; Atterbury, RJ and Connerton, IF (2011). Application of a bacteriophage cocktail to reduce Salmonella Typhimurium U288 contamination on pig skin. Int. J. Food Microbiol., 151: 157-163.
Huang, C; Virk, SM; Shi, J; Zhou, Y; Willias, SP; Morsy, MK; Abdelnabby, HE; Liu, J; Wang, X and Li, J (2018). Isolation, characterization, and application of bacteriophage LPSE1 against Salmonella enterica in ready to eat (RTE) foods. Front. Microbiol., 9: 1046.
Huff, WE; Huff, GR; Rath, NC; Balog, JM and Donoghue, AM (2002). Prevention of Escherichia coli infection in broiler chickens with a bacteriophage aerosol spray. Poult. Sci., 81: 1486-1491.
Shibiny, AE (2016). Bio-control of E. coli and Salmonella in foods using bacteriophage to improve food safety. World J. Dairy Food Sci., 11: 150-155.
Su, LH; Chiu, CH; Chu, C and Ou, JT (2004). Antimicrobial resistance in nontyphoid Salmonella serotypes: a global challenge. Clin. Infect. Dis., 39: 546-551.
Sukumaran, AT; Nannapaneni, R; Kiess, A and Sharma, CS (2016). Reduction of Salmonella on chicken breast
fillets stored under aerobic or modified atmosphere packaging by the application of lytic bacteriophage preparation SalmoFreshTM. Poult. Sci., 95: 668-675.
Thung, TY; Krishanthi Jayarukshi Kumari Premarathne, JM; San Chang, W; Loo, YY; Chin, YZ; Kuan, CH; Tan, CW; Basri, DF; Jasimah Wan Mohamed Radzi, CW and Radu, S (2017). Use of a lytic bacteriophage to control Salmonella Enteritidis in retail food. LWT. 78: 222-225.
Ye, J; Kostrzynska, M; Dunfield, K and Warriner, K (2010). Control of Salmonella on sprouting mung bean and alfalfa seeds by using a biocontrol preparation based on antagonistic bacteria and lytic bacteriophages. J. Food Prot., 73: 9-17. doi: 10.4315/0362-028X-73.1.9.
Zhang, X; Niu, YD; Nan, Y; Stanford, K; Holley, R; McAllister, T and Narvaez-Bravo, C (2019). SalmoFresh effectiveness in controlling Salmonella on romaine lettuce, mung bean sprouts and seeds. Int. J. Food Microbiol., 305: 108250. doi: 10.1016/j.ijfoodmicro.2019.108250.