Advances in Clinical and Experimental Medicine

Title abbreviation: Adv Clin Exp Med
JCR Impact Factor (IF) – 2.1 (5-Year IF – 2.0)
Journal Citation Indicator (JCI) (2023) – 0.4
Scopus CiteScore – 3.7 (CiteScore Tracker 3.8)
Index Copernicus  – 171.00; MNiSW – 70 pts

ISSN 1899–5276 (print)
ISSN 2451-2680 (online)
Periodicity – monthly

Download original text (EN)

Advances in Clinical and Experimental Medicine

2017, vol. 26, nr 3, May-June, p. 483–490

doi: 10.17219/acem/62245

Publication type: original article

Language: English

Download citation:

  • BIBTEX (JabRef, Mendeley)
  • RIS (Papers, Reference Manager, RefWorks, Zotero)

Anti-enterococcal activities of pentacyclic triterpenes

Dorota Wojnicz1,A,B,C,D, Dorota Tichaczek-Goska1,B,C,D, Kamila Korzekwa2,B, Marta Kicia1,B, Andrzej Hendrich1,E,F

1 Department of Biology and Medical Parasitology, Wroclaw Medical University, Poland

2 Institute of Genetics and Microbiology, University of Wrocław, Poland

Abstract

Background. Asiatic (AA) and ursolic (UA) acids are widely studied phytochemicals, but their antimicrobial properties are still poorly understood. Therefore our research has focused on their activity against uropathogenic Enterococcus faecalis strains.
Objectives. The aim of this research was to determine the influence of AA and UA on the growth, cell morphology, virulence factors and biofilm formation by E. faecalis strains.
Material and Methods. AA and UA were purchased from Sigma-Aldrich. E. faecalis strains were isolated from the urine samples of patients with urinary tract infections. The strains were checked for the presence of virulence genes using the PCR method. Their antimicrobial susceptibility was performed using the disc diffusion method. The MICs of triterpenes were determined using the broth microdilution method. The hydrophobicity of cells was established by salt aggregation test. Lipase and lecithinase activities were determined by using an agar medium containing egg yolk emulsion. DNase agar was used for the detection of DNase synthesis. Hemolytic activity was established using a sheep-blood agar. Todd-Hewitt agar medium containing gelatin was used for determination of gelatinase activity. The anti-biofilm activity of asiatic acid and ursolic acid was tested on polystyrene microtiter plates. It was examined using time-kill and biofilm assays.
Results. Reduction of growth and enzyme synthesis after exposure of E. faecalis to the acids was observed. None of the acids changed the hydrophobicity of bacteria. Stronger anti-biofilm activity was observed when the bacteria were incubated with AA. Thus, reduction of both the survival and the virulence factors will make bacteria less infectious.
Conclusion. Based on the results obtained, we can assume that the triterpenes investigated should be considered natural components of a human diet rather than as antibacterial agents used on their own.

Key words

Enterococcus faecalis, pentacyclic triterpenes, asiatic acid, ursolic acid, antimicrobial agents

References (39)

  1. Jäger S, Trojan H, Kopp T, Laszczyk MN, Scheffler A. Pentacyclic triterpene distribution in various plants – rich sources for a new group of multi-potent plant extracts. Molecules. 2009;14:2016–2031.
  2. Somova LO, Nadar A, Rammanan P, Shode FO. Cardiovascular, antihyperlipidemic and antioxidant effect of oleanolic acid and ursolic acid in experimental hypertension. Phytomedicine. 2003;10:115–121.
  3. Sultana T, Rashid MA, Ali MA, Mahmood SF. Hepatoprotective and antibacterial activity of ursolic acid extracted from Hedyotis corymbosa L. Bangladesh J Sci Ind Res. 2010;45:27–34.
  4. Krishnamurthy RG, Senut MC, Zemke D, et al. Asiatic acid, a pentacyclic triterpene from Centella asiatica, is neuroprotective in a mouse model of focal cerebral ischemia. J Neurosci Res. 2009;87:2541–2550.
  5. Shanmugam MK, Dai X, Kumar AP, Tan BK, Sethi G, Bishayee A. Ursolic acid in cancer prevention and treatment: Molecular targets, pharmacokinetics and clinical studies. Biochem Pharmacol. 2013;85:1579–1587.
  6. Tsai SJ, Yin MC. Antioxidative and anti-inflammatory protection of oleanolic acid and ursolic acid in PC12 cells. J Food Sci. 2008;73:174–178.
  7. Kurek A, Grudniak AM, Szwed M, et al. Oleanolic acid and ursolic acid affect peptidoglycan metabolism in Listeria monocytogenes. Anton Leeuw Int J G. 2010;97:61–68.
  8. Ren D, Zuo R, Gonzalez Barrios AF, Bedzyk LA, Eldridge GR, Pasmore ME, Wood TK: Differential gene expression for investigation of Escherichia coli biofilm inhibition by plant extract ursolic acid. Appl Environ Microbiol 2005, 71, 4022-4034.
  9. Zhou L, Ding Y, Chen W, Zhang P, Chen Y, Lv X. The in vitro study of ursolic acid and oleanolic acid inhibiting cariogenic microorganisms as well as biofilm. Oral Dis. 2013;19:494–500.
  10. Wojnicz D, Kicia M, Tichaczek-Goska D. Effect of asiatic and ursolic acids on morphology, hydrophobicity and adhesion of UPECs to uroepithelial cells. Folia Microbiol. 2013;58:245–252.
  11. Wojnicz D, Tichaczek-Goska D, Kicia M. Effect of asiatic and ursolic acids on growth and virulence factors of uropathogenic Escherichia coli strains. Turk J Biol. 2013;37:556–564.
  12. Wojnicz D, Tichaczek-Goska D, Kicia M. Pentacyclic triterpenes combined with ciprofloxacin help to eradicate the biofilm formed in vitro by Escherichia coli. Indian J Med Res. 2015;141:343–353.
  13. Rathnayake IU, Hargreaves M, Huygens F. Antibiotic resistance and virulence traits in clinical and environmental Enterococcus faecalis and Enterococcus faecium isolates. Syst Appl Microbiol. 2012;35:326–333.
  14. Zhu X, Wang Q, Zhang C, Cheung GS, Shen Y. Prevalence, phenotype, and genotype of Enterococcus faecalis isolated from saliva and root canals in patients with persistent apical periodontitis. J Endod. 2010;36:1950–1955.
  15. Clinical and Laboratory Standards Institute. Performance Standards for Antimicrobial Susceptibility Testing; Twenty-fourth Informational Supplement. CLSI Publication M100-S24, Vol. 34, No. 1. Wayne, PA, USA. Clinical and Laboratory Standards Institute, 2014.
  16. European Committee on Antimicrobial Susceptibility Testing (EUCAST). Breakpoint tables for interpretation of MICs and zone diameters, version 5.0, 2015.
  17. Cosentino S, Podda GS, Corda A, Fadda ME, Deplano M, Pisano MB. Molecular detection of virulence factors and antibiotic resistance pattern in clinical Enterococcus faecalis strains in Sardinia. J Prev Med Hyg. 2010;51:31–36.
  18. Creti R, Imperi M, Bertuccini L, et al. Survey for virulence determinants among Enterococcus faecalis isolated from different sources. J Med Microbiol. 2004;53:13–20.
  19. Lindahl M, Faris A, Wadström T, Hjertén S. A new test based on ‘salting out’ to measure relative surface hydrophobicity of bacterial cells. Biochim Biophys Acta. 1981;677:471–476.
  20. Furumura MT, Figueiredo PMS, Carbonell GV, Darini AL, Yano T. Virulence-associated characteristics of Enterococcus faecalis strains isolated from clinical sources. Braz J Microbiol. 2006;37:230–236.
  21. Harrigan WF. Laboratory Methods in Food Microbiology. San Diego, CA: Academic Press. 1998.
  22. Russell FM, Biribo SSN, Selvaraj G, et al. As a bacterial culture medium, citrated sheep blood agar is a practical alternative to citrated human blood agar in laboratories of developing countries. J Clin Microbiol. 2006;44:3346–3351.
  23. Zeng J, Teng F, Murray BE. Gelatinase is important for translocation of Enterococcus faecalis across polarized human enterocyte-like T84 cells. Infect Immun. 2005;73:1606–1612.
  24. Szewczyk EM. Bacteriological diagnostics. Warsaw, Poland: PWN; 2011.
  25. O’Toole GA, Kolter R. Flagellar and twitching motility are necessary for Pseudomonas aeruginosa biofilm development. Mol Microbiol. 1998;30:295–304.
  26. Stepanovic S, Vukovic D, Hola V, et al. Quantification of biofilm in microtiter plates: overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci. Acta Path Micro Im B. 2007;115:891–899.
  27. Di Bonaventura G, Spedicato I, D’Antonio D, Robuffo I, Piccolomini R. Biofilm formation by Stenotrophomonas maltophilia modulation by quinolones, trimetoprim-sulfamethoxazole, and ceftazidime. Antimicrob Agents Chemother. 2004;48:151–160.
  28. Al-Talib H, Zuraina N, Kamarudin B, Yean CY. Genotypic variations of virulent genes in Enterococcus faecium and Enterococcus faecalis isolated from three hospitals in Malaysia. Adv Clin Exp Med. 2015;24:121–127.
  29. Taemchuay D, Rukkwamsuk T, Sakpuaram T, Ruangwises N. Antibacterial activity of crude extracts of Centella asiatica against Staphylococcus aureus in bovine mastitis. Kasetsart Veterinarians. 2009;19:119–128.
  30. Liu WH, Liu TC, Mong MC. Antibacterial effects and action modes of asiatic acid. Biomedicine. 2015;5:22–29.
  31. Horiuchi K, Shiota S, Hatano T, Yoshida T, Kuroda T, Tsuchiya T. Antimicrobial activity of oleanolic acid from Salvia officinalis and related compounds on vancomycin-resistant enterococci (VRE). Biol Pharm Bull. 2007;30:1147–1149.
  32. Fontanay S, Grare M, Mayer J, Finance C, Duval RE. Ursolic, oleanolic and betulinic acids: Antibacterial spectra and selectivity indexes. J Ethnopharmacol. 2008;120:272–276.
  33. Ford M. Medical Microbiology. Oxford, UK: Oxford University Press; 2014.
  34. Kim MJ, Kim CS, Park JY, et al. Antimicrobial effects of ursolic acid against mutans streptococci isolated from Koreans. Int J Oral Biol. 2011;36:7–11.
  35. Broniatowski M, Flasiński M, Zięba K, Miśkowiec P. Interactions of pentacyclic triterpene acids with cardiolipins and related phosphatidylglycerols in model systems. Biochim Biophys Acta. 2014;1838:2530–2538.
  36. Ramirez-Arcos S, Liao M, Marthaler S, Rigden M, Dillon JA. Enterococcus faecalis divIVA: An essential gene involved in cell division, cell growth and chromosome segregation. Microbiology. 2005;151:1381–1393.
  37. Tsikrikonis G, Maniatis AN, Labrou M, et al. Differences in biofilm formation and virulence factors between clinical and fecal enterococcal isolates of human and animal origin. Microb Pathog. 2012;52:336–343.
  38. Schneewinda O, Missiakas D. Sec-secretion and sortase-mediated anchoring of proteins in gram-postive bacteria. Biochim Biophys Acta. 2014;1843:1687–1697.
  39. Garo E, Eldridge GR, Goering MG, et al. Asiatic acid and corosolic acid enhance the susceptibility of Pseudomonas aeruginosa biofilms to tobramycin. Antimicrob Agents Chemother. 2007;51:1813–1817.