High efficacy of a characterized lytic bacteriophage in combination with thyme essential oil against multidrug-resistant Staphylococcus aureus in chicken products

Document Type : Full paper (Original article)

Authors

1 Department of Food Control, Faculty of Veterinary Medicine, Zagazig University, 44511, Zagazig, Egypt (current address)

2 Zoonoses department, faculty of veterinary medicine, zagazig university, zagazig, Egypt

Abstract

Background: The emergence and spread of methicillin-resistant Staphylococcus aureus (MRSA) in the food industry have led to using alternative natural bioagents for controlling S. aureus in food. Aims: The current work aimed to isolate and characterize a lytic phage specific to S. aureus and evaluate its efficacy with thyme essential oil for controlling S. aureus growth in chicken fillets. Methods: Twenty S. aureus strains previously isolated from ready-to-eat chicken products were tested for antimicrobial susceptibility and used for phage isolation. Results: All S. aureus strains were multidrug-resistant (MDR). The isolated phage (vB_SauM_CP9) belonged to the family Myoviridae and maintained its stability at pH (4-9) and temperature (30-70°C). The phage showed lytic activity on ten S. aureus strains and had a burst size (228 PFU/infected cell), latent period (45 min), and rise period (15 min). A combination of S. aureus phage multiplicity of infection (MOI) 10 + thyme oil 1% caused a higher significant reduction in S. aureus growth (87.22%) in artificially inoculated chicken fillets than individual treatment with bacteriophage or thyme essential oil. Conclusion: To our knowledge, this is the first report to evaluate the efficacy of bacteriophage and thyme oil for controlling the growth of MDR S. aureus in chicken products and recommending application of S. aureus phage and thyme oil combination in the food industry to achieve food safety goals and consumer protection as well as mitigate the antimicrobial resistance crisis.

Keywords

Main Subjects

References

Ackermann, HW and Prangishvili, D (2012). Prokaryote viruses studied by electron microscopy. Arch. Virol., 157: 1843-1849.
Adriaenssens, E and Brister, JR (2017). How to name and classify your phage: an informal guide. Viruses. 9:70. doi:10.3390/v9040070.
Alves, DR; Gaudion, A; Bean, JE; Perez Esteban, P; Arnot, TC; Harper, DR; Kot, W; Hansen, LH; Enright, MC; Tobias, A and Jenkins, AT (2014). Combined use of bacteriophage K and a novel bacteriophage to reduce Staphylococcus aureus biofilm formation. Appl. Environ. Microbiol., 80: 6694-6703.
Anžlovar, S; Baričevič, D; Ambrožič Avguštin, J and Dolenc Koce, J (2014). Essential oil of common thyme as a natural antimicrobial food additive. Food Technol. Biotechnol., 52: 263-268.
Apata, D (2009). Antibiotic resistance in poultry. Int. J. Poult. Sci., 8: 404-408.
Atterbury, RJ; Connerton, PL; Dodd, CE; Rees, CE and Connerton, IF (2003). Application of host-specific bacteriophages to the surface of chicken skin leads to a reduction in recovery of Campylobacter jejuni. Appl. Environ. Microbiol., 69: 6302-6306.
Bajpai, VK; Baek, KH and Kang, SC (2012). Control of Salmonella in foods by using essential oils: a review. Food Res. Int., 45: 722-734.
Bakkali, F; Averbeck, S; Averbeck, D and Idaomar, M (2008). Biological effects of essential oils – a review. Food Chem. Toxicol., 46: 446-475.
Bassolé, IHN and Juliani, HR (2012). Essential oils in combination and their antimicrobial properties. Molecules. 17: 3989-4006.
Burt, S (2004). Essential oils: their antibacterial properties and potential applications in foods–a review. Int. J. Food Microbiol., 94: 223-253.
Chang, HC; Chen, CR and Lin, JW (2005). Isolation and characterization of novel giant Stenotrophomonas maltophilia phage phiSMA5. Appl. Environ. Microbiol., 71: 1387-1393.
Clinical and Laboratory Standards Institute (CLSI) (2019). Performance standards for antimicrobial susceptibility testing. 29th Edn., CLSI Supplement M100. Wayne, PA: CLSI.
Crago, B; Ferrato, C; Drews, S; Svenson, L; Tyrrell, G and Louie, M (2012). Prevalence of Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) in food samples associated with foodborne illness in Alberta, Canada from 2007 to 2010. Food Microbiol.,32: 202-205.
El Haddad, L; Abdallah, NB; Plante, PL; Dumaresq, J; Katsarava, R; Labrie, S; Corbeil, J; St-Gelais, D and Moineau, S (2014). Improving the safety of Staphylococcus aureus polyvalent phages by their production on a Staphylococcus xylosus strain. PLoS One. 9: e102600. https://doi: 10.1371/journal.pone.0102600.
Fan, Y; Li, S; Deng, B and Zhao, Y (2015). Prevalence and relevance analysis of multidrug-resistant Staphylococcus aureus of meat, poultry and human origin. Indian J. Anim. Res., 49: 86-90.
Fischetti, VA (2010). Bacteriophage endolysins: a novel anti-infective to control Gram-positive pathogens. Int. J. Med. Microbiol., 300: 357-362.
Ganaie, MY; Qureshi, S; Kashoo, Z; Wani, SA; Hussain, MI; Kumar, R; Maqbool, R; Sikander, P; Banday, MS; Malla, WA; Mondal, P and Khan, RIN (2018). Isolation and characterization of two lytic bacteriophages against Staphylococcus aureus from India: Newer therapeutic agents against bovine mastitis. Vet. Res. Commun., 42: 289-295.
Gharieb, RMA; Saad, MF; Mohamed, AS and Tartor, YH (2020). Characterization of two novel lytic bacteriophages for reducing biofilms of zoonotic multidrug-resistant Staphylococcus aureus and controlling their growth in milk. LWT-Food Sci. Technol., 124:109145. https://doi. org/10.1016/j.lwt.2020.109145.
Ghosh, A; Ricke, SC; Almeida, G and Gibson, KE (2016). Combined application of essential oil compounds and bacteriophage to inhibit growth of Staphylococcus aureus in vitro. Current Microbiol., 72: 426-435.
Goode, D; Allen, V and Barrow, P (2003). Reduction of experimental Salmonella and Campylobacter con-tamination of chicken skin by application of lytic bacteriophages. Appl. Environ. Microbiol., 69: 5032-5036.
Gutierrez, D; Vandenheuvel, D; Martinez, B; Rodriguez, A; Lavigne, R and Garcia, P (2015). Two phages, phiIPLA-RODI and phiIPLA-C1C, lyse mono- and dual-species staphylococcal biofilms. Appl. Environ. Microbiol., 81: 3336-3348.
Hammerum, AM; Lester, CH and Heuer, OE (2010). Antimicrobial-resistant enterococci in animals and meat: a human health hazard? Foodborne Pathog. Dis., 7: 1137-1146.
Ibrahim, HM; El Sabagh, RA; El-Roos, A; Nahla, A and Abd El Fattah, H (2016). Antimicrobial effect of some essential oils on Staphylococcus aureus in minced meat. Benha Vet. Med. J., 30: 183-191.
ICMSF (International Commission of Microbiological Specification for Foods) (1996). Microorganisms in Food. Their significance and methods of enumeration. 3rd Edn., Canada, University of Toronto.
Jończyk, E; Kłak, M; Międzybrodzki, R and Górski, A (2011). The influence of external factors on bacteriophages. Folia Microbiol., 56: 191-200.
Jun, JW; Kim, JH; Shin, SP; Han, JE; Chai, JY and Park, SC (2013). Characterization and complete genome sequence of the Shigella bacteriophage pSf-1. Res. Microbiol., 164: 979-986.
Kazi, M and Annapure, US (2016). Bacteriophage biocontrol of foodborne pathogens. J. Food Sci. Technol.,53: 1355-1362.
Kittler, S; Wittmann, J; Mengden, RALP; Klein, G; Rohde, C and Lehnherr, H (2017). The use of bacteriophages as One-Health approach to reduce multidrug-resistant bacteria. Sustain. Chem. Pharm., 5: 80-83.
Kropinski, AM; Prangishvili, D and Lavigne, R (2009). Position paper: the creation of a rational scheme for the nomenclature of viruses of bacteria and archaea. Environ. Microbiol., 11: 2775-2777.
Kvachadze, L; Balarjishvili, N; Meskhi, T; Tevdoradze, E; Skhirtladze, N; Pataridze, T; Adamia, R; Topuria, T; Kutter, E; Rohde, C and Kutateladze, M (2011). Evaluation of lytic activity of staphylococcal bacteriophage Sb-1 against freshly isolated clinical pathogens. Microb. Biotechnol., 4: 643-650.
Kwan, T; Liu, J; DuBow, M; Gros, P and Pelletier, J (2005). The complete genomes and proteomes of 27 Staphylococcus aureus bacteriophages. Proc. Natl. Acad. Sci. U. S. A., 102: 5174-5179.
Le Loir, Y; Baron, F and Gautier, M (2003). Staphylococcus aureus and food poisoning. Genet. Mol. Res., 2: 63-76.
Li, L and Zhang, Z (2014). Isolation and characterization of a virulent bacteriophage SPW specific for Staphylococcus aureus isolated from bovine mastitis of lactating dairy cattle. Mol. Biol. Rep., 41: 5829-5838.
Lone, A; Anany, H; Hakeem, M; Aguis, L; Avdjian, AC; Bouget, M; Atashi, A; Brovko, L; Rochefort, D and Griffiths, MW (2016). Development of prototypes of bioactive packaging materials based on immobilized bacteriophages for control of growth of bacterial pathogens in foods. Int. J. Food Microbiol., 217: 49-58.
Lu, Z; Breidt, JrF; Fleming, H; Altermann, E and Klaenhammer, T (2003). Isolation and characterization of a Lactobacillus plantarum bacteriophage, ΦJL-1, from a cucumber fermentation. Int. J. Food Microbiol., 84: 225-235.
Magiorakos, AP; Srinivasan, A; Carey, R; Carmeli, Y; Falagas, M; Giske, C; Harbarth, S; Hindler, JF; Kahlmeter, G; Olsson-Liljequist, B; Paterson, DL; Rice, LB; Stelling, J; Struelens, MJ; Vatopoulos, A; Weber, JT and Monnet, DL (2012). Multidrug-resistant, extensively drug-resistant and pandrug-resistant bacteria: an international expert proposal for interim standard definitions for acquired resistance. Clin. Microbiol. Infect., 18: 268-281.
Marek, A; Pyzik, E; Stepien-Pysniak, D; Urban-Chmiel, R and Nowaczek, A (2019). Characterization of bacterio-phages and their carriage in Staphylococcus aureus isolated from broilers in Poland. Br. Poult. Sci., 60: 373-380.
Moon, SH; Waite-Cusic, J and Huang, E (2020). Control of Salmonella in chicken meat using a combination of a commercial bacteriophage and plant-based essential oils. Food Control. 110: 106984. https://doi.org/10.1016/j. foodcont.2019.106984.
Nazzaro, F; Fratianni, F; De Martino, L; Coppola, R and De Feo, V (2013). Effect of essential oils on pathogenic bacteria. Pharmaceuticals. 6: 1451-1474.
Pajunen, M; Kiljunen, S and Skurnik, M (2000). Bacteriophage φYeO3-12, specific for Yersinia enterocolitica serotype O: 3, is related to coliphages T3 and T7. J. Bacteriol., 182: 5114-5120.
Pecarski, D; Ketin, S; Omerovic, I; Mirkovic, M; Jugovic, Z and Biocanin, R (2016). Chemical compositions and antimicrobial activities of oregano and thyme essential oils. Bulg. Chem. Commun., 48: 678-683.
Pual, S; Bezbarauh, RL; Roy, MK and Ghosh, AC (1997). Multiple antibiotic resistance index and its reversion in Pseudomonas aeruginosa. Lett. Appl. Microbiol., 24: 169-171.
Salem, AM; Zakaria, EM and Abd El-Raheem, KA (2017). Efficiency of some essential oils in control of methicillin resistant Staphylococcus aureus (MRSA) in Minced Beef. Benha Vet. Med. J., 32: 177-183.
Sambrook, JW and Russell, D (2001). Molecular cloning: a
laboratory manual. 3rd Edn., NY, USA, Cold Spring Harbor Press.
Schnabel, EL and Jones, AL (2001). Isolation and characterization of five Erwinia amylovora bacteriophages and assessment of phage resistance in strains of Erwinia amylovora. Appl. Environ. Microbiol., 67: 59-64.
Sulakvelidze, A (2013). Using lytic bacteriophages to eliminate or significantly reduce contamination of food by foodborne bacterial pathogens. J. Sci. Food Agric., 93: 3137-3146.
Synnott, AJ; Kuang, Y; Kurimoto, M; Yamamichi, K; Iwano, H and Tanji, Y (2009). Isolation from sewage influent and characterization of novel Staphylococcus aureus bacteriophages with wide host ranges and potent lytic capabilities. Appl. Environ. Microbiol., 75: 4483-4490.
Takemura-Uchiyama, I; Uchiyama, J; Kato, SI; Ujihara, T; Daibata, M and Matsuzaki, S (2014). Genomic and phylogenetic traits of Staphylococcus phages S25-3 and S25-4 (family Myoviridae, genus Twort-like viruses). Ann. Microbiol., 64: 1453-1456.
Teramoto, H; Salaheen, S and Biswas, D (2016). Contamination of post-harvest poultry products with multidrug resistant Staphylococcus aureus in Maryland-Washington DC metro area. Food Control. 65: 132-135.
Tey, BT; Ooi, ST; Yong, KC; Ng, MYT; Ling, TC and Tan, WS (2009). Production of fusion m13 phage bearing the di-sulphide constrained peptide sequence (C-WSFFSNI-C) that interacts with hepatitis B core antigen. Afr. J. Biotechnol., 8: 268-273.
Tongnuanchan, P and Benjakul, S (2014). Essential oils: Extraction, bioactivities, and their uses for food preservation. J. Food Sci., 79: 1231-1249.
Vandersteegen, K; Mattheus, W; Ceyssens, PJ; Bilocq, F; De Vos, D; Pirnay, JP; Noben, JP; Merabishvili, M; Lipinska, U; Hermans, K and Lavigne, R (2011). Microbiological and molecular assessment of bacteriophage ISP for the control of Staphylococcus aureus. PloS One. 6: e24418. doi: 10.1371/journal.pone. 0024418.
Yoon, H; Yun, J; Lim, JA; Roh, E; Jung, KS; Chang, Y; Ryu, S and Heu, S (2013). Characterization and genomic analysis of two Staphylococcus aureus bacteriophages isolated from poultry/livestock farms. J. Gen. Virol., 94: 2569-2576.

Files

Share

How to cite

Statistics