Original Article

Effects of nisin on the expression of virulence genes of methicillin-resistant/sensitive Staphylococcus aureus


Background and Objectives: Few studies have considered potential benefits of probiotic bacteria and their derivatives on human and animal health. Nisin is an antimicrobial agent that is produced by lactobacilli and served as a preservative in foods. This study aims to investigate whether nisin suppresses or decreases the genes involved in the pathogenicity of methicillin-susceptible and methicillin-resistant Staphylococcus aureus (MSSA and MRSA).
Materials and Methods: MSSA and MRSA strains were cultured at the ¼, ½, and 1 × minimum inhibitory concentration (MIC) of nisin. Next, RNA extraction was performed at the mid-exponential stage of growth, and cDNA was synthesized. The expression of virulence factors was measured by qPCR, and the data were analyzed by the ΔΔCt formula.
Results: Depending on the incubation times and the Lactobacillus species, the MIC of nisin on MRSA and MSSA observed in 800 and 1600 mg/l, respectively. The qPCR assay showed the expression level of the sea, agrA, and spa genes decreased and the level of the sae gene increased at the sub-MIC of nisin, and no antagonism was observed. Concerning MRSA, the maximum downregulation rate was observed in the sea gene (up to 5.9 folds) while in MSSA, the maximum downregulation rate was noticed in the agrA gene (up to 10 folds).
Conclusion: Due to the high inhibitory effect of the sub-MIC of nisin on the expression of virulence factor genes in MRSA and MSSA, this compound could potentially reduce the virulence of S. aureus.

1. Wilson J, Guy R, Elgohari S, Sheridan E, Davies J, Lamagni T, et al. Trends in sources of meticillin-resistant Staphylococcus aureus (MRSA) bacteraemia: data from the national mandatory surveillance of MRSA bacteraemia in England, 2006–2009. J Hosp Infect 2011; 79:211-217.
2. Chaibenjawong P, Foster SJ. Desiccation tolerance in Staphylococcus aureus. Arch Microbiol 2011; 193: 125-135.
3. Hennekinne J-A, De Buyser M-L, Dragacci S. Staphylococcus aureus and its food poisoning toxins: characterization and outbreak investigation. FEMS Microbiol Rev 2012; 36: 815-836.
4. Le Loir Y, Baron F, Gautier M. Staphylococcus aureus and food poisoning. Genet Mol Res 2003; 2: 63-76.
5. Schlecht LM, Peters BM, Krom BP, Freiberg JA, Hänsch GM, Filler SG, et al. Systemic Staphylococcus aureus infection mediated by Candida albicans hyphal invasion of mucosal tissue. Microbiology (Reading) 2015; 161: 168-181.
6. Yan X, Wang B, Tao X, Hu Q, Cui Z, Zhang J, et al. Characterization of Staphylococcus aureus strains associated with food poisoning in Shenzhen, China. Appl Environ Microbiol 2012; 78: 6637-6642.
7. Levy SB, Marshall B. Antibacterial resistance worldwide: causes, challenges and responses. Nat Med 2004; 10(12 Suppl): S122-9.
8. Livermore DM. Bacterial resistance: origins, epidemiology, and impact. Clin Infect Dis 2003; 36(Suppl 1): S11-S23.
9. Sengupta S, Chattopadhyay MK, Grossart H-P. The multifaceted roles of antibiotics and antibiotic resistance in nature. Front Microbiol 2013; 4: 47.
10. Di Giannatale E, Prencipe V, Tonelli A, Marfoglia C, Migliorati G. Characterisation of Staphylococcus aureus strains isolated from food for human consumption. Vet Ital 2011; 47: 165-173.
11. Fetsch A, Contzen M, Hartelt K, Kleiser A, Maassen S, Rau J, et al. Staphylococcus aureus food-poisoning outbreak associated with the consumption of ice-cream. Int J Food Microbiol 2014; 187: 1-6.
12. Lee J-Y, Kim Y-S, Shin D-H. Antimicrobial synergistic effect of linolenic acid and monoglyceride against Bacillus cereus and Staphylococcus aureus. J Agric Food Chem 2002; 50: 2193-2199.
13. Ragle BE, Wardenburg JB. Anti-alpha-hemolysin monoclonal antibodies mediate protection against Staphylococcus aureus pneumonia. Infect Immun 2009; 77: 2712-2718.
14. Le KY, Otto M. Quorum-sensing regulation in staphylococci—an overview. Front Microbiol 2015; 6: 1174.
15. Thompson TA, Brown PD. Association between the agr locus and the presence of virulence genes and pathogenesis in Staphylococcus aureus using a Caenorhabditis elegans model. Int J Infect Dis 2017; 54: 72-76.
16. Liu Q, Yeo WS, Bae T. The SaeRS two‐component system of Staphylococcus aureus. Genes (Basel) 2016; 7: 81.
17. Bennett S, Ben Said L, Lacasse P, Malouin F, Fliss I. Susceptibility to nisin, bactofencin, pediocin and reuterin of multidrug resistant Staphylococcus aureus, Streptococcus dysgalactiae and Streptococcus uberis causing bovine mastitis. Antibiotics 2021; 10: 1418.
18. Hasper HE, de Kruijff B, Breukink E. Assembly and stability of nisin− lipid II pores. Biochemistry 2004;43:11567-11575.
19. Kindrachuk J, Jenssen H, Elliott M, Nijnik A, Magrangeas-Janot L, Pasupuleti M, et al. Manipulation of innate immunity by a bacterial secreted peptide: lantibiotic nisin Z is selectively immunomodulatory. Innate Immun 2013; 19: 315-327.
20. Naghmouchi K, Le Lay C, Baah J, Drider D. Antibiotic and antimicrobial peptide combinations: synergistic inhibition of Pseudomonas fluorescens and antibiotic-resistant variants. Res Microbiol 2012; 163: 101-108.
21. Thomas LV, Ingram RE, Bevis HE, Davies EA, Milne CF, Delves-Broughton J. Effective use of nisin to control Bacillus and Clostridium spoilage of a pasteurized mashed potato product. J Food Prot 2002; 65: 1580-1585.
22. Dosler S, Gerceker AA. In vitro activities of antimicrobial cationic peptides; melittin and nisin, alone or in combination with antibiotics against Gram-positive bacteria. J Chemother 2012; 24: 137-143.
23.Mataraci E, Dosler S. In vitro activities of antibiotics and antimicrobial cationic peptides alone and in combination against methicillin-resistant Staphylococcus aureus biofilms. Antimicrob Agents Chemother 2012; 56: 6366-6371.
24. Ruijter JM, Ramakers C, Hoogaars WM, Karlen Y, Bakker O, Van den Hoff MJ, et al. Amplification efficiency: linking baseline and bias in the analysis of quantitative PCR data. Nucleic Acids Res 2009; 37(6): e45.
25. Naffa RG, Bdour SM, Migdadi HM, Shehabi AA. Enterotoxicity and genetic variation among clinical Staphylococcus aureus isolates in Jordan. J Med Microbiol 2006; 55: 183-187.
26. Wiedemann I, Breukink E, van Kraaij C, Kuipers OP, Bierbaum G, de Kruijff B, et al. Specific binding of nisin to the peptidoglycan precursor lipid II combines pore formation and inhibition of cell wall biosynthesis for potent antibiotic activity. J Biol Chem 2001; 276: 1772-1779.
27. Kim T-S, Hur J-W, Yu M-A, Cheigh C-I, Kim K-N, Hwang J-K, et al. Antagonism of Helicobacter pylori by bacteriocins of lactic acid bacteria. J Food Prot 2003; 66: 3-12.
28. Józefiak D, Kierończyk B, Juśkiewicz J, Zduńczyk Z, Rawski M, Długosz J, et al. Dietary nisin modulates the gastrointestinal microbial ecology and enhances growth performance of the broiler chickens. PloS one 2013; 8: e85347.
29. De Kwaadsteniet M, Doeschate K, Dicks LMT. Nisin F in the treatment of respiratory tract infections caused by Staphylococcus aureus. Lett Appl Microbiol 2009; 48: 65-70.
30. Aranha C, Gupta S, Reddy K. Contraceptive efficacy of antimicrobial peptide Nisin: in vitro and in vivo studies. Contraception 2004; 69: 333-338.
31. Gupta SM, Aranha CC, Reddy K. Evaluation of developmental toxicity of microbicide Nisin in rats. Food Chem Toxicol 2008; 46: 598-603.
32. Cao LT, Wu JQ, Xie F, Hu SH, Mo Y. Efficacy of nisin in treatment of clinical mastitis in lactating dairy cows. J Dairy Sci 2007; 90: 3980-3985.
33. Wu J, Hu S, Cao L. Therapeutic effect of nisin Z on subclinical mastitis in lactating cows. Antimicrob Agents Chemother 2007; 51: 3131-3135.
34. Brumfitt W, Salton MRJ, Hamilton-Miller JMT. Nisin, alone and combined with peptidoglycan-modulating antibiotics: activity against methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci. J Antimicrob Chemother 2002; 50: 731-734.
35. Giacometti A, Cirioni O, Barchiesi F, Scalise G. In-vitro activity and killing effect of polycationic peptides on methicillin-resistant Staphylococcus aureus and interactions with clinically used antibiotics. Diagn Microbiol Infect Dis 2000; 38: 115-118.
36. Dosler S, Gerceker AA. In vitro activities of nisin alone or in combination with vancomycin and ciprofloxacin against methicillin-resistant and methicillin-susceptible Staphylococcus aureus strains. Chemotherapy 2011; 57: 511-516.
37. Zhao X, Meng R, Shi C, Liu Z, Huang Y, Zhao Z, et al. Analysis of the gene expression profile of Staphylococcus aureus treated with nisin. Food Control 2016; 59: 499-506.
38. Balamurugan P, Praveen Krishna V, Bharath D, Lavanya R, Vairaprakash P, Adline Princy. S. Staphylococcus aureus quorum regulator SarA targeted compound, 2-[(Methylamino) methyl] phenol inhibits biofilm and down-regulates virulence genes. Front Microbiol 2017; 8:1290.
39. Azizkhani M, Misaghi A, Basti AA, Gandomi H, Hosseini H. Effects of Zataria multiflora Boiss. essential oil on growth and gene expression of enterotoxins A, C and E in Staphylococcus aureus ATCC 29213. Int J Food Microbiol 2013; 163: 159-165.
40. Nygaard TK, Pallister KB, Ruzevich P, Griffith S, Vuong C, Voyich JM. SaeR binds a consensus sequence within virulence gene promoters to advance USA300 pathogenesis. J Infect Dis 2010; 201: 241-254.
41. You Y-O, Choi N-Y, Kang S-Y, Kim K-J. Antibacterial activity of Rhus javanica against methicillin-resistant Staphylococcus aureus. Evid Based Complement Alternat Med 2013; 2013: 549207.
42. Azizkhani M, Akhondzadeh Basti A, Tooryan F. Comparing inhibitory potential of Eugenia caryophyllus and Origanum compactum against the growth and gene expression of enterotoxins in Staphylococcus aureus ATCC 29213. Iran J Vet Med 2017; 11: 299-311.
43. Merino N, Toledo-Arana A, Vergara-Irigaray M, Valle J, Solano C, Calvo E, et al. Protein A-mediated multicellular behavior in Staphylococcus aureus. J Bacteriol 2009; 191: 832-843.
44. Mun S-H, Kong R, Seo Y-S, Zhou T, Kang O-H, Shin D-W, et al. Subinhibitory concentrations of punicalagin reduces expression of virulence-related exoproteins by Staphylococcus aureus. FEMS Microbiol lett 2016; 363: fnw253.
IssueVol 14 No 6 (2022) QRcode
SectionOriginal Article(s)
DOI https://doi.org/10.18502/ijm.v14i6.11262
Nisin; Methicillin-resistant Staphylococcus aureus; Methicillin-sensitive Staphylococcus aureus; Virulence factor; Food-borne

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How to Cite
Ramezani M, Rezazadeh Zarandi E, Zainodini N, Bahramabadi R, Assar S. Effects of nisin on the expression of virulence genes of methicillin-resistant/sensitive Staphylococcus aureus. Iran J Microbiol. 2022;14(6):874-880.