Evaluation of antimicrobial resistance and immune evasion cluster genes in clinical methicillin-resistant Staphylococcus aureus (MRSA) isolates from Khuzestan Province, Iran

Document Type : Original Research

Authors
1 Department of Biology, Faculty of Science, Shahid Chamran University of Ahvaz, Ahvaz, Iran
2 Department of Biology, Faculty of Science, Shahid Chamran University of Ahvaz
Abstract


Aims: Methicillin-resistant Staphylococcus aureus (MRSA) is recognized as an important health problem worldwide. To counteract the human innate immunity, S. aureus produces a number of immune evasion cluster (IEC) including staphylokinase (SAK), staphylococcal enterotoxin P (SEP), staphylococcal enterotoxin A (SEA), staphylococcal complement inhibitor (SCIN), and chemotaxis inhibitory protein (CHIP) encoded by sak, sep, sea, scn, and chp genes, respectively. These genes are carried by β-hamolysin-converting bacteriophages. The present study was conducted to determine the IEC phage types and antibiotic resistance patterns in 145 clinical MRSA isolates from Khuzestan Province, Iran.

Methods: All the isolates were investigated by disc diffusion method and PCR assay of sak, sep, sea, scn, and chp genes.

Findings: The assessment of antibiotic resistance showed the highest rate of resistance towards penicillin (97.25%), followed by methicillin (95.8%), ceftazidime (81.4%), erythromycin (71.8%), clindamycin (61.4%), ciprofloxacin (60.7%), gentamycin (56%), imipenem (56.55%), and vancomycin (0%), respectively. Also, the frequency of IEC types was as follows: type A (4.8%), type B (9%), type C (13.1%), type D (12.4%), type E (27.6%), type F (1.4%), type G (0.7%), and type H (6.9%). On the other hand, 24.1% of the isolates did not show any of the IEC types.

Conclusion: The findings showed that IEC-carrying bacteriophages are highly prevalent among MRSA strains, resulting in the adaptation and counteraction of bacteria to the human immune system. Therefore, understanding the role of IEC in the virulence of bacteria can improve our knowledge about the evolution, vaccination, and treatment of S. aureus infection.

Keywords


1. Baptistão LG, Silva NC, Bonsaglia EC, Rossi BF, Castilho IG, Fernandes Junior A, et al. Presence of Immune Evasion Cluster and Molecular Typing of Methicillin-Susceptible Staphylococcus aureus Isolated from Food Handlers. J Food Prot. 2016; 79(4):682-6.
2. Liang BS, Huang YM, Chen YS, Dong H, Mai JL, Xie YQ, et al. Antimicrobial resistance and prevalence of CvfB, SEK and SEQ genes among Staphylococcus aureus isolates from paediatric patients with bloodstream infections. Exp Ther Med. 2017; 14(5):5143-5148.
3. Hau SJ, Sun J, Davies PR, Frana TS, Nicholson TL. Comparative Prevalence of Immune Evasion Complex Genes Associated with β-Hemolysin Converting Bacteriophages in MRSA ST5 Isolates from Swine, Swine Facilities, Humans with Swine Contact, and Humans with No Swine Contact. PLoS One. 2015; 10(11):e0142832.
4. McCarthy AJ1, Witney AA, Lindsay JA. Staphylococcus aureus temperate bacteriophage: carriage and horizontal gene transfer is lineage associated. Front Cell Infect Microbiol. 2012; 2:6.
5. Foster TJ. Immune evasion by staphylococci. Nat Rev Microbiol. 2005; 3(12):948-58.
6. Yu F, Li T, Huang X, Xie J, Xu Y, Tu J, et al. Virulence gene profiling and molecular characterization of hospital‑acquired Staphylococcus aureus isolates associated with bloodstream infection. Diagn Microbiol Infect Dis. 2012; 74(4):363-8.
7. Jongerius I, von Köckritz-Blickwede M, Horsburgh MJ, Ruyken M, Nizet V, Rooijakkers SH. Staphylococcus aureus virulence is enhanced by secreted factors that block innate immune defenses. J Innate Immun. 2012; 4(3):301-11.
8. van Wamel WJ, Rooijakkers SH, Ruyken M, van Kessel KP, van Strijp JA. The innate immune modulators staphylococcal complement inhibitor and chemotaxis inhibitory protein of Staphylococcus aureus are located on beta-hemolysin-converting bacteriophages. J Bacteriol. 2006; 188(4):1310-5.
9. Verkaik NJ, Benard M, Boelens HA, de Vogel CP, Nouwen JL, Verbrugh HA, et al. Immune evasion cluster-positive bacteriophages are highly prevalent among human Staphylococcus aureus strains, but they are not essential in the first stages of nasal colonization. Clin Microbiol Infect. 2011; 17(3):343-8.
10. Jongerius I, Köhl J, Pandey MK, Ruyken M, van Kessel KP, van Strijp JA, et al. Staphylococcal complement evasion by various convertase-blocking molecules. J Exp Med. 2007; 204(10):2461-71.
11. Kraushaar B, Hammerl JA, Kienöl M, Heinig ML, Sperling N, Dinh Thanh M, et al. Acquisition of virulence factors in livestock-associated MRSA: Lysogenic conversion of CC398 strains by virulence gene-containing phages. Sci Rep. 2017; 7(1):2004.
12. de Jong NWM, Vrieling M, Garcia BL, Koop G, Brettmann M, Aerts PC, et al. Identification of a staphylococcal complement inhibitor with broad host specificity in equid Staphylococcus aureus strains. J Biol Chem. 2018; 293(12):4468-4477.
13. CLSI, Performance Standards for Antimicrobial Susceptibility Testing; Twenty-Second Informational Supplement. 2017; 37:58-64.
14. Emaneini M, Bigverdi R, Kalantar D, Soroush S, Jabalameli F, Noorazar Khoshgnab B, et al. Distribution of genes encoding tetracycline resistance and aminoglycoside modifying enzymes in Staphylococcus aureus strains isolated from a burn center. Ann Burns Fire Disasters 2013; 26(2):76-80.
15. Ahmadrajabi R, Layegh-Khavidaki S, Kalantar-Neyestanaki D, Fasihi Y. Molecular analysis of immune evasion cluster (IEC) genes and intercellular adhesion gene cluster (ICA) among methicillin-resistant and methicillin-sensitive isolates of Staphylococcus aureus. J Prev Med Hyg. 2017; 58(4): E308-E314.
16. Price LB, Stegger M, Hasman H, Aziz M, Larsen J, Andersen PS, et al. Staphylococcus aureus CC398: host adaptation and emergence of methicillin resistance in livestock. MBio. 2012; 3(1): e00305-11.
17. Stefani S, Chung DR, Lindsay JA, Friedrich AW, Kearns AM, Westh H, et al. Meticillin-resistant Staphylococcus aureus (MRSA): global epidemiology and harmonisation of typing methods. Int J Antimicrob Agents. 2012; 39(4):273-82.
18. Hudson LO, Murphy CR, Spratt BG, Enright MC, Elkins K, Nguyen C, et al. Diversity of methicillin-resistant Staphylococcus aureus (MRSA) strains isolated from inpatients of 30 hospitals in Orange County, California. PLoS One. 2013; 8(4):e62117.
19. Dinges MM, Orwin PM, Schlievert PM. Exotoxins of Staphylococcus aureus. Clin Microbiol Rev. 2000; 13(1): 16 34.
20. You Y, Song L, Nonyane BAS, Price LB, Silbergeld EK. Genomic differences between nasal Staphylococcus aureus from hog slaughterhouse workers and their communities. PLoS One. 2018; 13(3):e0193820.
21. Zadoks RN, van Leeuwen WB, Kreft D, Fox LK, Barkema HW, Schukken YH, et al. Comparison of Staphylococcus aureus isolates from bovine and human skin, milking equipment, and bovine milk by phage typing, pulsed-field gel electrophoresis, and binary typing. J Clin Microbiol. 2002; 40(11):3894–3902.
22. Goerke C, Wirtz C, Fluckiger U, Wolz C. Extensive phage dynamics in Staphylococcus aureus contributes to adaptation to the human host during infection. Mol Microbiol 2006; 61(6):1673-85.
23. Sadeghi J, Mansouri S. Molecular characterization and antibiotic resistance of clinical isolates of methicillin-resistant Staphylococcus aureus obtained from Southeast of Iran (Kerman). APMIS 2014; 122(5):405-11.
24. Ohadian Moghadam S, Pourmand MR, Mahmoudi M, Sadighian H. Molecular characterization of methicillin-resistant Staphylococcus aureus: characterization of major clones and emergence of epidemic clones of sequence type (ST) 36 and ST 121 in Tehran, Iran. FEMS Microbiol Lett. 2015; 362(8):fnv043.
25. Saderi H, Emadi B, Owlia P. Phenotypic and genotypic study of macrolide, lincosamide and streptogramin B (MLSB) resistance in clinical isolates of Staphylococcus aureus in Tehran, Iran. Med Sci Monit. 2011;17(2):BR48-53.