Evaluation of the antibacterial sensitivity properties of mannitol salt agar

Authors

Keywords:

antimicrobial susceptibility testing; disk diffusion; high-salt medium; mannitol salt agar; Mueller-Hinton agar; Staphylococcus aureus; vancomycin

Abstract

Background: Staphylococcus spp. are major pathogens causing diverse clinical infections. Evaluating their phenotypic characteristics and antimicrobial response across different culture media is essential for accurate diagnosis.

Objective: To assess vancomycin susceptibility testing performance of Mannitol Salt Agar (MSA) compared with Mueller–Hinton Agar (MHA) against Staphylococcus spp.

Methods: Thirty clinical specimens (urine, wound, ear swabs) were collected. Isolates were identified using standard microbiological methods including MSA for colony color, blood agar for hemolysis, and VITEK-2 confirmation. Vancomycin susceptibility was evaluated on both media. Statistical analysis used t-test, chi-square, and Pearson correlation (p < 0.05).

Results: Staphylococcus spp. were predominantly isolated from urine (60%; p = 0.007). On MSA, 40% showed yellow colonies and 60% pinkish-white (p = 0.27). Hemolysis patterns were 46.7% beta and 53.3% alpha/gamma (p > 0.05). Vancomycin produced significantly larger inhibition zones on MSA (25.3 ± 6.1 mm) than MHA (17.5 ± 5 mm; p < 0.0001). Strong correlation existed between colony color and hemolysis pattern (r = 0.873, p < 0.001).

Conclusions: Vancomycin showed significantly larger inhibition zones on MSA compared to MHA, indicating that high salt content and mannitol alter antibiotic diffusion and/or bacterial phenotypic expression. Larger zones on MSA represent greater apparent susceptibility rather than enhanced resistance detection. Therefore, MSA cannot replace MHA for standardized antimicrobial susceptibility testing, as altered medium composition leads to misleading susceptibility profiles.

Downloads

Download data is not yet available.

References

1. Gherardi G. Staphylococcus aureus Infection: Pathogenesis and Antimicrobial Resistance [Internet]. Int J Mol Sci. 2023;24(9):8182. DOI: 10.3390/ijms24098182

2. Al-Mebairik NF, El-kersh TA, Al-Sheikh YA, Marie MA. A review of virulence factors, pathogenesis, and antibiotic resistance in Staphylococcus aureus [Internet]. Rev Med Microbiol. 2016;27(2):51-56. DOI: 10.1097/MRM.0000000000000067

3. Nogueira JO, da Silva JD, de Pascoa Junior JG, Pereira TC, Bezerra CC. Staphylococcus aureus resistente em unidade de terapia intensiva [Internet]. Res Soc Dev. 2023;12(6):e7312. DOI: 10.33448/rsd-v12i6.42006

4. Nikolic P, Mudgil P. The Cell Wall, Cell Membrane and Virulence Factors of Staphylococcus aureus and Their Role in Antibiotic Resistance [Internet]. Microorganisms. 2023;11(2):259. DOI: 10.3390/microorganisms11020259

5. Kumar S, Singh S, Dhanjal DS. Pathogenesis and Antibiotic Resistance of Staphylococcus aureus. In: Arora G, Sajid A, Kalia VC, editors. Microbial Pathogenesis: Strategies for Diagnosis and Therapy [Internet]. Singapore: Springer; 2020. p. 1-20. DOI: 10.1007/978-981-15-1695-5_7

6. Mukherjee R, Priyadarshini A, Pandey RP, Raj VS. Antimicrobial Resistance in Staphylococcus aureus [Internet]. In: Islam Aqib A, Rodríguez-Morales AJ. Insights Into Drug Resistance in Staphylococcus aureus. London: IntechOpen; 2021. DOI: 10.5772/intechopen.96888

7. Monaco M, Araujo FP, Cruciani M, Coccia EM, Pantosti A. Worldwide Epidemiology and Antibiotic Resistance of Staphylococcus aureus [Internet]. Curr Top Microbiol Immunol. 2016;409:21-56. DOI: 10.1007/82_2016_3

8. Stefani S, Goglio A. Methicillin-resistant Staphylococcus aureus: related infections and antibiotic resistance [Internet]. Int J Infect Dis. 2010; (Suppl 4):S19-22. DOI: 10.1016/j.ijid.2010.05.003

9. Hu Q, Peng H, Rao X. Molecular events for promotion of vancomycin resistance in vancomycin intermediate Staphylococcus aureus [Internet]. Front Microbiol. 2016;7:1601. DOI: 10.3389/fmicb.2016.01601

10. Selim S, Faried OA, Almuhayawi MS. Incidence of Vancomycin-Resistant Staphylococcus aureus Strains among Patients with Urinary Tract Infections [Internet]. Antibiotics. 2022;11(3):408. DOI: 10.3390/antibiotics11030408

11. Patel H, Rawat S. A genetic regulatory see-saw of biofilm and virulence in MRSA pathogenesis [Internet]. Front Microbiol. 2023;14:1204428. DOI: 10.3389/fmicb.2023.1204428

12. Tasneem U, Majid M, Mehmood K. Co-occurrence of antibiotic resistance and virulence genes in Methicillin Resistant Staphylococcus aureus (MRSA) isolates from Pakistan [Internet]. Afr Health Sci. 2022;22(2):567-77. DOI: 10.4314/ahs.v22i1.57

13. Mesrati I, Saidani M, Jemili M. Virulence determinants, biofilm production and antimicrobial susceptibility in Staphylococcus aureus causing device-associated infections [Internet]. Int J Antimicrob Agents. 2018;52(3):322-7. DOI: 10.1016/j.ijantimicag.2018.05.004

14. Alkuraythi DM. Virulence Factors and Pathogenicity of Staphylococcus aureus [Internet]. In: Bustos J, Valdés JJ, Hamdan A, Rodríguez AJ. Advances and Perspectives of Infections Caused by Staphylococcus aureus. London: IntechOpen; 2024. DOI: 10.5772/intechopen.1006633

15. Graf AC, Leonard A, Schauble M. Virulence Factors Produced by Staphylococcus aureus Biofilms Have a Moonlighting Function Contributing to Biofilm Integrity [Internet]. Mol Cell Proteomics. 2019;18(5):1036-52. DOI: 10.1074/mcp.RA118.001120

16. Ballah FM, Islam MS, Rana M. Virulence Determinants and Methicillin Resistance in Biofilm-Forming Staphylococcus aureus from Food Sources [Internet]. Antibiotics. 2022;11(11):1666. DOI: 10.3390/antibiotics11111666

17. Humphries R, Bobenchik AM, Hindler JA, Schuetz AN. Overview of changes to the CLSI performance standards for antimicrobial susceptibility testing, M100 [Internet]. J Clin Microbiol. 2021;59(3):e00213-21. DOI: 10.1128/JCM.00213-21

18. Yousefi M, Fallah F, Arshadi M, Pourmand MR, Hashemi A, Pourmand G. Identification of tigecycline- and vancomycin-resistant Staphylococcus aureus strains in urinary tract infection patients [Internet]. New Microbes New Infect. 2017;19:8-12. DOI: 10.1016/j.nmni.2017.05.009

19. McGuinness WA, Malachowa N, DeLeo FR. Vancomycin Resistance in Staphylococcus aureus [Internet]. Yale J Biol Med. 2017 [access: 13/04/2026];90(2):269-281. Available from: https://pmc.ncbi.nlm.nih.gov/articles/PMC5482303/

20. Tong SYC, Davis JS, Eichenberger E, Holland TL, Fowler VG. Staphylococcus aureus infections: epidemiology, pathophysiology, clinical manifestations, and management [Internet]. Clin Microbiol Rev. 2023;36(3):e00013-23. DOI: 10.1128/cmr.00134-14

21. Cernohorska L. Antibiotic resistance and biofilm formation in Staphylococcus aureus strains isolated from urine [Internet]. Klin Mikrobiol Infekc Lek. 2010 [access: 13/04/2026];16(5):196-198. Available from: https://pubmed.ncbi.nlm.nih.gov/21243597/

22. Wang X, Zhan P, Zhang Q, Li R, Fan H. Staphylococcus aureus acquires resistance to vancomycin via CXCL10 [Internet]. Int Immunopharmacol. 2024;132:111780. DOI: 10.1016/j.intimp.2024.111780

23. Sahle B, Merid Y. Prevalence and antibiotic resistance of Staphylococcus aureus in wound infections [Internet]. J Infect Dev Ctries. 2024;18(10):1530-8. DOI: 10.3855/jidc.19023

24. Stojanovic-Radic Z, Dimitrijevic M, Stankovic N, Aleksic A, Pejcic M. Frequency of isolation and antibiotic resistance patterns of bacterial isolates from wound infections [Internet]. Biol Nyssana. 2016;7:151-158. DOI: 10.5281/zenodo.200414

25. Vandenesch F, Lina G, Henry T. Staphylococcus aureus hemolysins, bi-component leukocidins, and cytolytic peptides: A redundant arsenal of membrane-damaging virulence factors? [Internet]. Front Cell Infect Microbiol. 2012;2:12. DOI: 10.3389/fcimb.2012.00012 Available from: https://www.frontiersin.org/articles/10.3389/fcimb.2012.00012/full

26. de Albuquerque VV, Flores VC, Zeron RM, Godoi BB, Eulalio Filho WM, dos Santos BA. Study of vancomycin resistance among Staphylococcus aureus [Internet]. Amadeus Int Multidiscip J. 2019;3(6):31-37. DOI: 10.14295/aimj.v3i6.58

27. Olayinka BO, Olayinka AT, Onaolapo JA, Olurinola PF. Patterns of resistance to vancomycin and other agents in staphylococcal isolates [Internet]. Afr J Clin Exp Microbiol. 2005;6(1):21-27. DOI: 10.4314/ajcem.v6i1.7395

28. Pal M, Kerorsa GB, Marami LM, Kandi V. Epidemiology, pathogenicity, animal infections, antibiotic resistance, public health significance and economic impact of Staphylococcus aureus: a review [Internet]. Am J Public Health Res. 2020;8(1):14-21. DOI: 10.12691/ajphr-8-1-3

Downloads

Published

2026-05-01

How to Cite

1.
AL-Khikani F, Abd-alhussein MS, Kadhim Madloul K. Evaluation of the antibacterial sensitivity properties of mannitol salt agar. Rev. cuba. med. mil [Internet]. 2026 May 1 [cited 2026 May 3];55(2):e026077196. Available from: https://revmedmilitar.sld.cu/index.php/mil/article/view/77196