Determination of the effect of glucose, sucrose and sodium chloride addition in different culture media on biofilm formation of methicillin resistant Staphylococcus aureus

Yıl 2022, Cilt: 4 Sayı: 2, 152 - 157, 29.03.2022

Ali Doğan Dursun , Samet Uçak , Orhan Yavuz , Mediha Nur Zafer Yurt , Behiye Büşra Taşbaşı , Elif Esma Acar , Veli Cengiz Özalp , Mert Sudağıdan

https://doi.org/10.38053/acmj.1037458

Cited By: 1

Öz

Aim: Staphylococcus aureus is the most clinically important bacterium among Staphylococci, colonizing 15-36% of the entire population. Biofilm formation is an important virulence factor of S. aureus. Treatment of biofilm-associated S. aureus infections is difficult. This study aimed to investigate the effects of glucose, sucrose, and sodium chloride (NaCl) addition to seven different media on biofilm formation capacity of methicillin resistant S. aureus (MRSA) strains.
Material and Method: Biochemical and molecular methods (spa, nuc, coa, and mecA PCR) were used to identify S. aureus strains. Cefoxitin resistance was determined by the agar disc diffusion method. Biofilm formation of the strains was investigated in 7 different media (Tryptone soya broth (TSB), TSB+1% sucrose, TSB+1% glucose, TSB+4% NaCl, Brain Heart Infusion broth (BHI), BHI+1% glucose, and BHI+4% NaCl) using the microplate test. The growth of strains in 7 different media was determined at 600 nm, and then 96-well microplates were stained with crystal violet and their biofilm formation abilities were determined by measuring absorbance values ​​at 590 nm.
Results: In this study, 53 strains containing spa, nuc, coa, and mecA genes were identified as MRSA with resistance to cefoxitin. When biofilm formation was examined in seven different media using the microplate test, the biofilm formation ability of MRSA strains increased significantly with glucose and sucrose addition to TSB and BHI (P<0.05), while the addition of 4% NaCl decreased the biofilm formation (P<0.05). When the media were compared, it was determined that BHI increased bacterial growth and biofilm formation compared to TSB.
Conclusion: It was concluded that the biofilm formation abilities of MRSA strains increased especially in the presence of glucose. The results showed that biofilm production capacity is affected by environmental factors, especially nutrient content in used media.

Anahtar Kelimeler

MRSA, Biofilm, Microplate test

Kaynakça

• Botelho-Nevers E, Gagnaire J, Verhoeven PO, et al. Decolonization of Staphylococcus aureus carriage. Med Mal Infect 2017; 47: 305-10.
• Horn J, Stelzner K, Rudel T, Fraunholz M. Inside job: Staphylococcus aureus host-pathogen interactions. Int J Med Microbiol 2018; 308: 607-24.
• Zainulabdeen SMS, Dakl AA. Pathogenicity and virulence factors in Staphylococcus aureus. Muthanna J Pure Sci 2021; 8: 109-19.
• Jin T, Mohammad M, Pullerits R, Ali A. Bacteria and host interplay in Staphylococcus aureus septic arthritis and sepsis. Pathogens 2021; 10: 1-25.
• Kanyo EC, Nowacki AS, Gordon SM, Shrestha NK. Comparison of mortality, stroke, and relapse for methicillin-resistant versus methicillin-susceptible Staphylococcus aureus infective endocarditis: a retrospective cohort study. Diagn Microbiol Infect Dis 2021; 100: 115395.
• Naicker PR, Karayem K, Hoek KGP, Harvey J, Wasserman E. Biofilm formation in invasive Staphylococcus aureus isolates is associated with the clonal lineage. Microb Pathog 2016; 90: 41-9.
• O’Neill E, Pozzi C, Houston P, et al. A novel Staphylococcus aureus biofilm phenotype mediated by the fibronectin-binding proteins, FnBPA and FnBPB. J Bacteriol 2008; 190: 3835-50.
• Pourzal F, Haghkhah M. Prevalence of biofilm associated genes in different isolates of Staphylococcus aureus. J Med Bacteriol 2020; 9: 9-15.
• Taj Y, Essa F, Aziz F, Kazmi SU. Study on biofilm-forming properties of clinical isolates of Staphylococcus aureus. J Infect Dev Ctries 2012; 6: 403-9.
• Oniciuc EA, Cerca N, Nicolau AI. Compositional analysis of biofilms formed by Staphylococcus aureus isolated from food sources. Front Microbiol 2016; 7: 2014-7.
• Otto M. Staphylococcal biofilms. Microbiol Spectrum 2018; 6: GPP3-0023-2018.
• Avila-Novoa MG, González-Gómez JP, Guerrero-Medina PJ, et al. Staphylococcus aureus and methicillin-resistant S. aureus (MRSA) strains isolated from dairy products: Relationship of ica-dependent/independent and components of biofilms produced in vitro. Int Dairy J 2021; 119: 105066.
• Azmi K, Qrei W, Abdeen Z. Screening of genes encoding adhesion factors and biofilm production in methicillin resistant strains of Staphylococcus aureus isolated from Palestinian patients. BMC Genomics 2019; 20: 1-12.
• Nyenje ME, Green E, Ndip RN. Evaluation of the effect of different growth media and temperature on the suitability of biofilm formation by Enterobacter cloacae strains isolated from food samples in South Africa. Molecules 2013; 18: 9582-93.
• Sugimoto S, Sato F, Miyakawa R, et al. Broad impact of extracellular DNA on biofilm formation by clinically isolated methicillin-resistant and -sensitive strains of Staphylococcus aureus. Sci Rep 2018; 8: 2254.
• Lade H, Park JH, Chung SH, et al. Biofilm Formation by Staphylococcus aureus clinical isolates is differentially affected by glucose and sodium chloride supplemented culture media. J Clin Med 2019; 8: 1853.
• Tang J, Chen J, Liu J, Zhang R, Yang R, Chen L. Effects of different cultivation conditions on Staphylococcus aureus biofilm formation and diversity of adhesin genes. J Food Safety 2012; 32: 210-8.
• Sudagidan M, Çavuşoğlu C, Bacakoğlu F. Investigation of the virulence genes in methicillin-resistant Staphylococcus aureus strains isolated from biomaterial surfaces. Mikrobiyol Bul 2008; 42: 29-39.
• Aires-De-Sousa M, Boye K, De Lencastre H, et al. High interlaboratory reproducibility of DNA sequence-based typing of bacteria in a multicenter study. J Clin Microbiol 2006; 44: 619-21.
• Sudagidan M, Aydin A. Screening virulence properties of Staphylococci isolated from meat and meat products. Wien Tierärztl Mschr 2009; 96: 128-34.
• Hookey J V, Richardson JF, Cookson BD. Molecular typing of Staphylococcus aureus based on PCR restriction fragment length polymorphism and DNA sequence analysis of the coagulase gene. J Clin Microbiol 1998; 36: 1083-9.
• Lem P, Spiegelman J, Toye B, Ramotar K. Direct detection of mecA, nuc and 16S rRNA genes in BacT/Alert blood culture bottles. Diagn Microbiol Infect Dis 2001; 41: 165-8.
• Clinical and Laboratory Standards Institute (CLSI). Performance Standards for Antimicrobial Susceptibility Testing Document M100. 30th ed. Pennsylvania, USA; 2020: 58-66.
• Stepanović S, Djukić V, Djordjević V, Djukić S. Influence of the incubation atmosphere on the production of biofilm by Staphylococci. Clin Microbiol Infect 2003; 9: 955-8.
• Sudagidan M, Erdem İ, Çavuşoğlu C, Çiftçioğlu M. Investigation of the surface properties of Staphylococcus epidermidis strains isolated from biomaterials. Mikrobiyol Bul 2010; 44: 93-103.
• Şahin R, Kaleli İ. Comparison of genotypic and phenotypic characteristics in biofilm production of Staphylococcus aureus isolates. Mikrobiyol Bul 2018; 52: 111-22.
• Lee S, Choi KH, Yoon Y. Effect of NaCl on biofilm formation of the isolate from Staphylococcus aureus outbreak linked to ham. Korean J Food Sci Anim Resour 2014; 34: 257-61.
• Knobloch JKM, Horstkotte MA, Rohde H, Mack D. Evaluation of different detection methods of biofilm formation in Staphylococcus aureus. Med Microbiol Immunol 2002; 191: 101-6.
• Waldrop R, McLaren A, Calara F, McLemore R. Biofilm growth has a threshold response to glucose in vitro. Clin Orthop Relat Res 2014; 472: 3305-10.
• Agarwal A, Jain A. Glucose & sodium chloride induced biofilm production & ica operon in clinical isolates of Staphylococci. Indian J Med Res 2013; 138: 262-6.
• McCarthy H, Rudkin JK, Black NS, Gallagher L, O’Neill E, O’Gara JP. Methicillin resistance and the biofilm phenotype in Staphylococcus aureus. Front Cell Infect Microbiol 2015; 5: 1-9.
• Singh A, Prakash P, Achra A, Singh G, Das A, Singh R. Standardization and classification of in vitro biofilm formation by clinical isolates of Staphylococcus aureus. J Glob Infect Dis 2017; 9: 93-101.
• Vázquez-Sánchez D, Habimana O, Holck A. Impact of food-related environmental factors on the adherence and biofilm formation of natural Staphylococcus aureus isolates. Curr Microbiol 2013; 66: 110-21.
• Xu H, Zou Y, Lee HY, Ahn J. Effect of NaCl on the biofilm formation by foodborne pathogens. J Food Sci 2010; 75: 580-5.
• Lee J-S, Bae Y-M, Lee S-Y, Lee S-Y. Biofilm formation of Staphylococcus aureus on various surfaces and their resistance to chlorine sanitizer. J Food Sci 2015; 80: M2279-86.
• Sudagidan M, Ozalp VC, Öztürk O, et al. Bacterial surface, biofilm and virulence properties of Listeria monocytogenes strains isolated from smoked salmon and fish contact surfaces. Food Biosci 2021; 101021.