In vitro activities of Traganum nudatum and Mentha pulegium extracts combined with amphotericin B against Candida albicans in production of hydrolytic enzymes

Document Type : Original Articles


1 Department of Antifungal Antibiotic, Physico-Chemical Synthesis, and Biological Activity, University of Tlemcen, Tlemcen, Algeria

2 University Center of Naâma, Naâma, Algeria

3 Department of Physiology, Physiopathology, and Biochemistry of Nutrition, University of Tlemcen, Tlemcen, Algeria


Background and Purpose: Candida albicans is an important microorganism in the normal flora of a healthy subject; however, it has an expedient pathogenic character that induces hydrolytic virulence. Regarding this, the present study aimed to find an in vitro alternative that could reduce the virulence of this yeast.
Materials and Methods: For the purpose of the study, the effect of amphotericin B (AmB) combined with the extract of Traganum nudatum (E1) or Mentha pulegium (E2) was evaluated against the hydrolytic activities of esterase, protease, and phospholipase. This effect was determined by calculating the minimum inhibitory concentration (MIC), used to adjust the extract/AmB mixtures in culture media.
Results: The evaluated Pz values, which corresponded to the different enzymatic activities, showed a decrease in the hydrolytic activities of C. albicans strains after the addition of E1/AmB and E2/AmB combinations at descending concentrations (lower than the obtained MICs).
Conclusion: Based on the findings, it would be possible to reduce the pathogenesis of this species without destabilizing the balance of the flora.


1. Zhang C, Wang W, Kong Q, Liu F, Chen J, Sang H. Characterization of a Candida albicans isolate from a recurrent cervical lymphadenitis patient. Mycologia. 2019; 111(6):942-52.
2. Ranjan A, Dongari-Bagtzoglou A. Tipping the Balance: C. albicans adaptation in polymicrobial environments. J Fungi. 2018; 4(3):112.
3. Kirchner FR, Littringer K, Altmeier S, Tran VD, Schönherr F, Lemberg C, et al. Persistence of Candida albicans in the oral mucosa induces a curbed inflammatory host response that is independent of immunosuppression. Front Immunol. 2019; 10:330.
4. Fidel PL Jr, Yano J, Esher SK, Noverr MC. Applying the host-microbe damage response framework to candida pathogenesis: current and prospective strategies to reduce damage. J Fungi. 2020; 6(1):35.
5. Montoya AM, Luna-Rodríguez CE, Gracia-Roblesa G, Rojas OC, Treviño-Rangel RJ, González GM. In vitro virulence determinants, comparative pathogenicity of Diutina (Candida) mesorugosa clinical isolates and literature review of the D. rugosa complex. Mycolgia. 2019; 111(3):395-407.
6. Pammi M, Holland L, Butler G, Gacser A, Bliss JM. Candida parapsilosis is a significant neonatal pathogen: a systematic review and meta-analysis. Pediatr Infect Dis J. 2013; 32(5):e206-16.
7. Schaller M, Borelli C, Korting HC, Hube B. Hydrolytic enzymes as virulence factors of Candida albicans. Mycoses. 2005; 48(6):365-77.
8. Ghannoum MA. Potential role of phospholipases in virulence and fungal pathogenesis. Clin Microbiol Rev. 2000; 13(1):122-43.
9. Kumar CG, Kumar SSJ, Menon T. Phospholipase and proteinase activities of clinical isolates of Candida from immunoco-mpromised patients. Mycopathologia. 2006; 161(4):213-8.
10. Dannaoui É. Resistance of Candida to antifungals: detection and mechanisms. French J Lab. 2013; 450:71-7.
11. Grela E, Piet M, Luchowski R, Grudzinski W, Paduch R, Gruszecki WI. Imaging of human cells exposed to an antifungal antibiotic amphotericin B reveals the mechanisms associated with the drug toxicity and cell defense. Sci Rep. 2018; 8(1):14067.
12. Bhat V, Sharma SM, Shetty V, Shastry CS, Rao V. Extracellular enzymes of Candida albicans and their role in development of denture stomatitis-a review. J Acquir Immune Defic Syndr. 2011; 2(1):26-30.
13. Badji B, Riba A, Mathieu F, Lebrihi A, Sabaou N. Antifungal activity of a strain of Actinomadura of Saharan origin on various pathogenic and toxinogenic fungi. J Med Mycol. 2005; 15(4):211-9.
14. Mouderas F, El Haci IA, Lahfa FB. Phytochemical profile, antioxidant and antimicrobial activities of Traganum nudatum Delile aerial parts organic extracts collected from Algerian Sahara’s flora. Orient Pharm Exp Med. 2019; 19(3):299-310.
15. Barhoumi M. Comparative study of chemical properties and composition of Algerian and Tunisian Mentha pulegium L. Am J Ethnomed. 2020; 7(1):16.
16. Samaranayake LP, Raeside JM, MacFarlane TW. Factors affecting the phospholipase activity of Candida species in vitro. Sabouraudia. 1984; 22(3):201-7.
17. Aoki S, Ito-Kuwa S, Nakamura Y, Masuhara T. Comparative pathogenicity of a wild-type strain and respiratory mutants of Candida albicans in mice. Zentralblatt Bakteriol. 1990; 273(3):332-43.
18. Slifkin M. Tween 80 opacity test responses of various Candida species. J Clin Microbiol. 2000; 38(12):4626-8.
19. Clinical and Laboratory Standard Institute. Reference method for broth dilution antifungal susceptibility testing of yeasts; approved standard. Document M27-A3. 3rd ed. Wayne, P.A: Clinical and Laboratory Standard Institute; 2008.
20. Seddiki SM, Boucherit-Otmani Z, Boucherit K, Kunkel D. Fungal infectivities of implanted catheters due to Candida sp. Biofilms formation and resistance. J Mycol Med. 2015; 25(2):130-5.
21. Silva-Rocha WP, de Brito Lemos VL, Ferreira MRA, Soares LA, Svidzisnki TI, Milan EP, et al. Effect of the crude extract of Eugenia uniflora in morphogenesis and secretion of hydrolytic enzymes in Candida albicans from the oral cavity of kidney transplant recipients. BMC Complement Altern Med. 2015; 15:6.
22. Price MF, Wilkinson ID, Gentry LO. Plate method for detection of phospholipase activity in Candida albicans. Sabouraudia. 1982; 20(1):7-14.
23. Cannon JP, Fiscella R, Pattharachayakul S, Garey KW, Alba FD, Piscitelli S, et al. Comparative toxicity and concentrations of intravitreal amphotericin B formulations in a rabbit model. Invest Ophthalmol Vis Sci. 2003; 44(5):2112-7.
24. Aouadhi C, Ghazghazi H, Hasnaoui B, Maaroufi A. Comparison of the antifungal activity of methanolic extracts from three plants collected from north-west Tunisia. Microbiol Hyg Alim. 2013; 25(73):9-14.
25. Calandra OM. Traitement des infections invasives à Candida et Aspergillus: quoi de neuf en 2003? Rev Med Suisse. 2003; 1:22901.
26. Guo N, Liu J, Wu X, Bi X, Meng R, Wang X, et al. Antifungal activity of thymol against clinical isolates of fluconazole-sensitive and –resistant Candida albicans. J Med Microbiol. 2009; 58(8):1074-9.
27. El Alama H, El Aissami A, Benmoussa A, Said AA, Arahou M, El Alaoui-Faris FE. Kinetics of essential oil-antifungal interactions. Bull R Soc Sci Liège. 2017; 86:67-77.
28. Rollenhagen C, Mamtani S, Ma D, Dixit R, Eszterhas S, Lee SA. The role of secretory pathways in Candida albicans pathogenesis. J Fungi. 2020; 6(1):26.
29. Santos AL, Braga-Silva LA. Aspartic protease inhibitors: effective drugs against the human fungal pathogen Candida albicans. Mini Rev Med Chem. 2013; 13(1):155-62.
30. Pandey N, Gupta MK, Tilak R. Extracellular hydrolytic enzyme activities of the different Candida spp. isolated from the blood of the Intensive Care Unit-admitted patients. J Lab Physicians. 2018; 10(4):392-6.
31. Ramos LD, Barbedo LS, Braga-Silva LA, dos Santos AL, Pinto MR, Sgarbi DB. Protease and phospholipase activities of
Candida spp. Isolated from cutaneous candidiasis. Rev Iberoam Micol. 2014; 32(2):122-5. -
32. Tsang CS, Chu FC, Leung WK, Jin LJ, Samaranayake LP, Siu SC. Phospholipase, proteinase and haemolytic activities of Candida albicans isolated from oral cavities of patients with type 2 diabetes mellitus. J Med Microbiol. 2007; 56(10):1393-8.
33. Price MF, Cawson RA. Phospholipase activity in Candida albicans. Sabouraudia. 1977; 15(2):179-85.
34. Cui J, Ren B, Tong Y, Dai H, Zhang L. Synergistic combinations of antifungals and anti-virulence agents to fight against Candida albicans. Virulence. 2015; 6(4):362-71.
35. Abaci O. Investigation of extracellular phospholipase and proteinase activities of Candida species isolated from individuals denture wearers and genotypic distribution of Candida albicans strains. Cur Microbiol. 2011; 62(4):1308-14.
36. Mierziak J, Kostyn K, Kulma A. Flavonoids as important molecules of plant interactions with the environment. Molecules. 2014; 19(10):16240-65.
37. Mohan V, Ballal M. Proteinase and phospholipase activity as virulence factors in Candida species isolated from blood. Rev Iberoam Micol. 2008; 25(4):208-10.
38. Budzyńska A, Sadowska B, Więckowska-Szakiel M, Różalska B. Enzymatic profile, adhesive and invasive properties of Candida albicans under the influence of selected plant essential oils. Acta Biochim Pol. 2014; 61(1):115-21.
Volume 6, Issue 3
September 2020
Pages 27-32
  • Receive Date: 19 June 2020
  • Revise Date: 26 August 2020
  • Accept Date: 12 September 2020
  • First Publish Date: 12 September 2020