Characterization of Biofilm Producing Staphylococcus epidermidis Strains Isolated from Patients and Healthy People

Document Type : Original Research

Authors
Department of Microbiology, Faculty of Biological Science and Technology, University of Isfahan, Isfahan, Iran
Abstract
Background: Staphylococcus epidermidis isolates are among the most important causes of nosocomial infections and could be classified as health threatening agents. This study aimed to determine the biofilm formation ability and clonal dissemination of S. epidermidis strains isolated from patients and healthy people in Isfahan during 2016 and 2017.

Materials & Methods: A total of 139 and 123 suspected colonies of S. epidermidis were collected from different clinical specimens and the arm of healthy people, respectively. The ability to form biofilm was determined using a combination of Congo-red agar (CRA) and microtiter plate (MTP) assays. The presence of genes involved in biofilm formation was also tested by the polymerase chain reaction (PCR) test. The susceptibility of all strains to 12 antibiotics was evaluated using the disk diffusion method according to the Clinical & Laboratory Standards Institute (CLSI) guidelines. Moreover, all biofilm-producing strains were typed using PhenePlate system as well as cassette chromosome mec (SCCmec) and accessory gene regulator (agr) locus typing method.

Findings: A total of 43 biofilm-producing S. epidermidis strains were identified among 107 and 123 confirmed strains isolated from hospitalized patients and healthy people, respectively; all of which were positive for aap gene, and the presence of ica operon was limited to 86 and 27% of the strains isolated from patients and healthy people, respectively. All the strains showed susceptibility to vancomycin, quinupristin-dalfopristin, and linezolid. Moreover, SCCmec Types III, IV, and V were detected among all methicillin-resistant S. epidermidis (MRSE) strains, and agr Type I was the most frequent one. Among all biofilm-positive strains, 3 common types (CTs) and 7 single types (STs) were determined; CT1 and CT2 were the most common types among the strains isolated from hospitalized patients and healthy people.

Conclusion: These findings indicated the presence and persistence of diverse clone types of biofilm-producing S. epidermidis strains with common types of PhP, agr, and SCCmec in the hospital and the community of Isfahan.

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1. Cheung GY, Otto M. Understanding the significance of Staphylococcus epidermidis bacteremia in babies and children. Current Opinion in Infectious Diseases. 2010;23(3):208.
2. Gomes F, Leite B, Teixeira P, Oliveira R. Strategies to control Staphylococcus epidermidis biofilms. In: Méndez-Vilas's A, editor. Science against microbial pathogens: communicating current research and technological advances Formatex Research Center; 2011. p. 843-52.
3. Fey PD, Olson ME. Current concepts in biofilm formation of Staphylococcus epidermidis. Future Microbiology. 2010;5(6):917-33.
4. Prasad S, Nayak N, Satpathy G, Nag H, Venkatesh P, Ramakrishnan S, et al. Molecular & phenotypic characterization of Staphylococcus epidermidis in implant related infections. Indian Journal of Medical Research. 2012;136(3):483.
5. O'Gara JP. ica and beyond: biofilm mechanisms and regulation in Staphylococcus epidermidis and Staphylococcus aureus. FEMS Microbiology Letters. 2007;270(2):179-88.
6. Chen M, Yu Q, Sun H. Novel strategies for the prevention and treatment of biofilm related infections. International Journal of Molecular Sciences. 2013;14(9):18488-501.
7. Līduma I, Tračevska T, Bērs U, Žileviča A. Phenotypic and genetic analysis of biofilm formation by Staphylococcus epidermidis. Medicina. 2012;48(6):45.
8. Rahimi F. Molecular characteristics of biofilm-producing methicillin-resistant Staphylococcus epidermidis isolates causing urinary tract infections. Archives of Clinical Infectious Diseases. 2018;13(6):e61704.
9. Xu L, Li H, Vuong C, Vadyvaloo V, Wang J, Yao Y, et al. Role of the luxS quorum-sensing system in biofilm formation and virulence of Staphylococcus epidermidis. Infection and Immunity. 2006;74(1):488-96.
10. Periasamy S, Joo H-S, Duong AC, Bach T-HL, Tan VY, Chatterjee SS, et al. How Staphylococcus aureus biofilms develop their characteristic structure. Proceedings of the National Academy of Sciences. 2012;109(4):1281-6.
11. Najar-Peerayeh S, Behmanesh M, Jazayeri Moghadas A. Staphylococcus epidermidis, conality and accessory gne regulator diversity in clinical isolates Archives of Clinical Infectious Diseases. 2018; 13(4):e62833.
12. Rahimi F, Katouli M, Karimi S. Biofilm production among methicillin resistant Staphylococcus aureus strains isolated from catheterized patients with urinary tract infection. Microbial Pathogenesis. 2016;98:69-76.
13. Stepanović S, Vuković D, Hola V, Bonaventura GD, Djukić S, Ćirković I, et al. Quantification of biofilm in microtiter plates: overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci. Apmis. 2007;115(8):891-9.
14. Clinical and Laboratory Standard Institute C. Performance standards for antimicrobial susceptibility testing, 26th informational supplement. Clinical and Laboratory Standard Institute, Wayne, Pa. 2016.
15. Rahimi F, Shokoohizadeh L. Characterization of methicillin resistant Staphylococcus aureus strains among inpatients and outpatients in a referral hospital in Tehran, Iran. Microbial Pathogenesis. 2016;97:89-93.
16. Arciola CR, Baldassarri L, Montanaro L. Presence of icaA and icaD genes and slime production in a collection of staphylococcal strains from catheter-associated infections. Journal of Clinical Microbiology. 2001;39(6):2151-6.
17. Conlon KM, Humphreys H, O'Gara JP. Inactivations of rsbU and sarA by IS256 represent novel mechanisms of biofilm phenotypic variation in Staphylococcus epidermidis. Journal of Bacteriology. 2004;186(18):6208-19.
18. Petrelli D, Zampaloni C, d’Ercole S, Prenna M, Ballarini P, Ripa S, et al. Analysis of different genetic traits and their association with biofilm formation in Staphylococcus epidermidis isolates from central venous catheter infections. European Journal of Clinical Microbiology and Infectious Diseases. 2006;25(12):773-781.
19. Rahimi F, Katouli M, Pourshafie MR. Characteristics of hospital-and community-acquired meticillin-resistant Staphylococcus aureus in Tehran, Iran. Journal of Medical Microbiology. 2014;63(Pt 6):796-804.
20. Lina G, Boutite F, Tristan A, Bes M, Etienne J, Vandenesch F. Bacterial competition for human nasal cavity colonization: role of Staphylococcal agr alleles. Applied and Environmental Microbiology. 2003;69(1):18-23.
21. Farajzadeh Sheikh A, Asareh Zadegan Dezfuli A, Navidifar T, Samei Fard S, Dehdashtian M. Association between biofilm formation, structure and antibiotic resistance in Staphylococcus epidermidis isolated from neonatal septicemia in southwest Iran. Infection and Drug Resistance. 2019;12:1771-82.
22. Solati SM, Tajbakhsh E, Khamesipour F, Gugnani HC. Prevalence of virulence genes of biofilm producing strains of Staphylococcus epidermidis isolated from clinical samples in Iran. AMB Express. 2015;5(1):47.
23. Zalipour M, Ebrahim-Saraie HS, Sarvari J, Khashei R. Detection of biofilm production capability and icaA/D genes among staphylococci isolates from Shiraz, Iran. Jundishapur Journal of Microbiology. 2016;9(12):e41431.
24. Ebrahimi A, Ghasemi M, Ghasemi B. Some virulence factors of staphylococci isolated from wound and skin infections in Shahrekord, IR Iran. Jundishapur Journal of Microbiology. 2014;7(4):e9225.
25. Deka N. Comparison of tissue culture plate method, tube method and Congo red agar method for the detection of biofilm formation by coagulase negative Staphylococcus isolated from non-clinical isolates. International Journal of Current Microbiology and Applied Sciences. 2014;3(10):810-5.
26. Gad GFM, El-Feky MA, El-Rehewy MS, Hassan MA, Abolella H, El-Baky RMA. Detection of icaA, icaD genes and biofilm production by Staphylococcus aureus and Staphylococcus epidermidis isolated from urinary tract catheterized patients. The Journal of Infection in Developing Countries. 2009;3(05):342-51.
27. Kord M, Ardebili A, Jamalan M, Jahanbakhsh R, Behnampour N, Ghaemi EA. Evaluation of biofilm formation and presence of ica genes in Staphylococcus epidermidis clinical isolates. Osong Public Health and Research Perspectives. 2018;9(4):160.
28. Galdbart J-O, Allignet J, Tung H-S, Rydèn C, El Solh N. Screening for Staphylococcus epidermidis markers discriminating between skin-flora strains and those responsible for infections of joint prostheses. Journal of Infectious Diseases. 2000;182(1):351-5.
29. Singhai M, Malik A, Shahid M, Malik A, Rawat V. Colonization of peripheral intravascular catheters with biofilm producing microbes: evaluation of risk factors. Nigerian Medical Journal. 2012;53(1):37.
30. Pinheiro L, Brito CI, Pereira VC, Oliveira Ad, Camargo CH, Cunha MdLRd. Reduced susceptibility to vancomycin and biofilm formation in methicillin-resistant Staphylococcus epidermidis isolated from blood cultures. Memorias do Instituto Oswaldo Cruz. 2014;109(7):871-8.
31. Wojtyczka R, Orlewska K, Kępa M, Idzik D, Dziedzic A, Mularz T, et al. Biofilm formation and antimicrobial susceptibility of Staphylococcus epidermidis strains from a hospital environment. International Journal of Environmental Research and Public Health. 2014;11(5):4619-33.
32. Du X, Zhu Y, Song Y, Li T, Luo T, Sun G, et al. Molecular analysis of Staphylococcus epidermidis strains isolated from community and hospital environments in China. PLoS One. 2013;8(5):e62742.
33. Saber H, Jasni AS, Jamaluddin TZMT, Ibrahim R. A review of Staphylococcal cassette chromosome mec (SCCmec) types in coagulase-negative staphylococci (CoNS) species. Malaysian Journal of Medical Sciences. 2017;24(5):7-18.
34. Shamansouri S, Karbasizade V, Khozaie M. Determining SCCmec types in Staphylococcus epidermidis isolated from clinical samples of Isfahan, Iran. Acta Medica Mediterranea. 2016;32:2107-2113.
35. Najar-Peerayeh S, Moghaddas AJ, Bakhshi B, Ghasemian A. Diversity of the SCCmec types among Staphylococcus epidermidis clinical isolates from intensive care unit patients. Asian Pacific Journal of Tropical Disease. 2016;6(2):133-5.
36. Carmody A, Otto M. Specificity grouping of the accessory gene regulator quorum-sensing system of Staphylococcus epidermidis is linked to infection. Archives of Microbiology. 2004;181(3):250-3.
37. Li M, Guan M, Jiang X, Yuan F, Xu M, Zhang W, et al. Genetic polymorphism of the accessory gene regulator (agr) locus in Staphylococcus epidermidis and its association with pathogenicity. Journal of Medical Microbiology. 2004;53(6):545-9.
38. Rahimi F. Characterization of resistance to aminoglycosides in methicillin-resistant Staphylococcus aureus strains isolated from a tertiary care hospital in Tehran, Iran. Jundishapur Journal of Microbiology. 2016;9(1):e29237.
39. Rahimi F, Bouzari M. Biochemical fingerprinting of methicillin-resistant Staphylococcus aureus isolated from sewage and hospital in Iran. Jundishapur Journal of Microbiology. 2015;8(7):e19760.
40. Rahimi F, Shafiei R. Characteristics of enterotoxin-producing methicillin-resistant Staphylococcus aureus strains isolated from meat in Tehran, Iran. Journal of Consumer Protection and Food Safety. 2019;14(4): 389-98.