Mikrobiol. Z. 2022; 84(5):21-29.
doi: https://doi.org/10.15407/microbiolj84.05.021

Synergistic Effect of Surfactants of Nocardia vaccinii IMV B-7405
and Essential Oils on Candida Genus Yeast

T.P. Pirog1,2, L.V. Kliuchka1, T.A. Shevchuk2, F.V. Muchnyk2

1National University of Food Technologies
68 Volodymyrska Str., Kyiv, 01601, Ukraine

2Zabolotny Institute of Microbiology and Virology, NAS of Ukraine
154 Akad. Zabolotny Str., Kyiv, 03143, Ukraine

The increase in the number of resistant strains of Candida genus representatives, capable of forming biofilms on various surfaces, stimulates the search for new, alternative methods of combating them, one of which is the use of compounds of natural origin, such as essential oils. However, at the same time, their concentration should be minimal, which is due to the ability of essential oils to cause severe damage of the human’s central nervous system and aspiration pneumonia. This leads to the necessity of searching for new methods to reduce the concentration of essential oils and at the same time to preserve their properties, in particular, by their use in a mixture with other antimicrobial agents, which can be microbial surfactants. Previously, it was found that the degree of yeast biofilm destruction under the action of Nocardia vaccinii IMV B-7405 surfactants dependson the nature of the growth substrate and is the highest in the presence of preparations synthesized on purified glycerol. Aim. To study the synergism of antifungal activity and the role in the destruction of biofilms of a mixture of Nocardia vaccinii IMV B-7405 surfactants synthesized on glycerol of different quality and essential oils. Methods. N. vaccinii IMV B-7405 was grown in a medium containing purified glycerol or waste from biodiesel production at a concentration of 2% (v/v) as carbon sources. The surfactants were extracted from the supernatant of cultural liquid by a modified Folch mixture. The antimicrobial activity of essential oils, surfactants, and their mixtures was determined by the index of the minimum inhibitory concentration. To assess the synergistic effect of a mixture of surfactants with essential oils, the fractional inhibitory concentration index was used. The degree of biofilm destruction (%) was determined as the difference between the cell adhesion in untreated and treated with surfactants, essential oil, or their mixture wells of the polystyrene microplates. Results. It was found that the surfactants synthesized by N. vaccinii IMV B-7405 on both purified glycerol and waste from biodiesel production showed synergistic antifungal activity in mixtures with cinnamon and lemongrass essential oils. Thus, the minimum inhibitory concentrations against Candida albicans D-6, Candida utilis BVS-65, and Candida tropicalis RE-2 of a mixture of surfactants synthesized on purified glycerol with cinnamon and lemongrass essential oils were 1.8—7.5 and 3.7— 15 μg/mL, respectively, and were lower than in the case of using surfactants (30-60 μg/mL), cinnamon or lemongrass essential oil (156—312 μg/mL) alone. The use of a mixture of surfactants obtained on waste from biodiesel production and cinnamon or lemongrass essential oils made it possible to reduce the minimum inhibitory concentrations of the latter against studied yeast test cultures by 14—56 times. At the same time, the index of fractional inhibitory concentration did not exceed 0.5, which indicates the synergism of the antifungal activity of the mixture of these compounds. The destruction of Candida yeast biofilms under the action of surfactants synthesized on both purified glycerol and waste from biodiesel production in a mixture with cinnamon or lemongrass essential oils reached 60—67 and 67—77%, respectively, which is an average of 25—35% higher compared to the use of each monopreparation separately. Conclusions. The results presented in this paper confirm the previously obtained data that N. vaccinii IMV B-7405 surfactants, synthesized on both traditional substrates and toxic industrial wastes, have antimicrobial and antiadhesive synergistic action with essential oils, which allows us to consider them as potential components of the so-called «antifungal locks» in the fight against of Candida genus representatives.

Keywords: Nocardia vaccinii IMV B-7405, surfactants, essential oils, yeast of Candida genus, synergism of antifungal activity, destruction of biofilms.

Full text

  1. Lewis JS, Wiederhold NP, Wickes BL, Patterson TF, Jorgensen JH. Rapid emergence of echinocandin resistance in Candida glabrata resulting in clinical and microbiologic failure. Antimicrob Agents Chemother. 2013; 57(9):4559—61. https://doi.org/10.1128/AAC.01144-13
  2. Perlin DS, Rautemaa-Richardson R, Alastruey-Izquierdo A. The global problem of antifungal resistance: prevalence, mechanisms, and management. Lancet Infect Dis. 2017; 17(12):383—92. https://doi.org/10.1016/S1473-3099(17)30316-X
  3. Arendrup MC, Patterson TF. Multidrug-Resistant Candida: epidemiology, molecular mechanisms, and treatment. J Infect Dis. 2017; 216(3):445—51. https://doi.org/10.1093/infdis/jix131
  4. Lohse MB, Gulati M, Johnson AD, Nobile CJ. Development and regulation of single- and multi-species Candida albicans biofilms. Nat Rev Microbiol. 2018; 16(1):19—31. https://doi.org/10.1038/nrmicro.2017.107
  5. Pohl CH. Recent Advances and opportunities in the study of Candida albicans polymicrobial biofi lms. Front Cell Infect Microbiol. 2022; 12:836379. https://doi.org/10.3389/fcimb.2022.836379
  6. Veilleux MP, Grenier D. Determination of the effects of cinnamon bark fractions on Candida albicans and oral epithelial cells. BMC Complement Altern Med. 2019; 19(1):303. https://doi.org/10.1186/s12906-019-2730-2
  7. Li Y, Shao X, Xu J, Wei Y, Xu F, Wang H. Effects and possible mechanism of tea tree oil against Botrytis cinerea and Penicillium expansum in vitro and in vivo test. Can J Microbiol. 2017; 63(3): 219—27. https://doi.org/10.1139/cjm-2016-0553
  8. Richards DB, Wang GS, Buchanan JA. Pediatric tea tree oil aspiration treated with surfactant in the emergency department. Pediatr Emerg Care. 2015; 31(4):279—80. https://doi.org/10.1097/PEC.0000000000000234
  9. Pirog TP, Kliuchka LV, Kliuchka IV, Shevchuk TA., Iutynska G.O. Synergism of antimicrobial and anti-adhesive activity of Nocardia vaccinii IMV B-7405 surfactants in a mixture with essential oils. Microbiol Z. 2020; 82(4):31—40. https://doi.org/10.15407/microbiolj82.04.031
  10. Pirog TP, Kliuchka LV, Shevchuk TA, Iutynska GO. Destruction of biofilms on silicone tubes under action of a mixture of Nocardia vaccinii IMV B-7405 surfactants with other biocides. Microbiol Z. 2021; 83(4):43—53. https://doi.org/10.15407/microbiolj83.04.043
  11. Pirog TP, Kliuchka IV, Kliuchka LV, Shevchuk TA, Iutynska GO. [Biofilm destruction in the presence of surfactants synthesized under different cultivation conditions of Nocardia vaccinii IMB B-7405]. Microbiol Z. 2019; 81(5):3−15. Ukrainian. https://doi.org/10.15407/microbiolj81.05.003
  12. Mazzola P, Jozala A, Lencastre-Novaes L, Moriel P, Vessoni-Penna T. Minimal inhibitory concentration (MIC) determination of disinfectant and/or sterilizing agents. Braz J Pharm Sci. 2009; 45(2):241—48. https://doi.org/10.1590/S1984-82502009000200008
  13. Hallander HO, Dornbusch K, Gezelius L, Jacobson K, Karlsson I. Synergism between aminoglycosides and cephalosporins with antipseudomonal activity: interaction index and killing curve method. Antimicrob Agents Chemother. 1982; 22(5):743—52. https://doi.org/10.1128/AAC.22.5.743
  14. Cavalheiro M, Teixeira MC. Candida biofilms: threats, challenges, and promising strategies. Front Med (Lausanne). 2018; 5:28. https://doi.org/10.3389/fmed.2018.00028
  15. de Barros PP, Rossoni RD, de Souza CM, Scorzoni L, Fenley JC, Junqueira JC. Candida biofilms: an update on developmental mechanisms and therapeutic challenges. Mycopathologia. 2020; 185(3):415—424. https://doi.org/10.1007/s11046-020-00445-w
  16. Jaroš P, Vrublevskaya M, Lokočová K, Michailidu J, Kolouchová I, Demnerová K. Boswellia serrata extract as an antibiofilm agent against Candida spp. microorganisms. 2022; 10(1):171. https://doi.org/10.3390/microorganisms10010171
  17. Shahina Z, El-Ganiny AM, Minion J, Whiteway M, Sultana T, Dahms TES. Cinnamomum zeylanicum bark essential oil induces cell wall remodelling and spindle defects in Candida albicans. Fungal Biol Biotechnol. 2018; 5:3. https://doi.org/10.1186/s40694-018-0046-5
  18. Pirog TP, Nikituk LV, Iutynska GO. [Biological properties of Nocardia vaccinii IMV B-7405 surfactants synthesized on by product of biodiesel production]. Microbiol Z. 2016; 78(5):12−20. Ukrainian. https://doi.org/10.15407/microbiolj78.05.012
  19. Feyaerts AF, Mathé L, Luyten W, De Graeve S, Van Dyck K, Broekx L, et al. Essential oils and their components are a class of antifungals with potent vapour-phase-mediated anti-Candida activity. Sci Rep. 2018; 8(1):3958. https://doi.org/10.1038/s41598-018-22395-6
  20. Bhattacharya R, Rolta R, Dev K, Sourirajan, A. Synergistic potential of essential oils with antibiotics to combat fungal pathogens: present status and future perspectives. Phytother Res. 2021; 35(11):6089—6100. https://doi.org/10.1002/ptr.7218
  21. Essid R, Hammami M, Gharbi D. Karkouch I, Hamouda TB , Elkahoui S, et al. Antifungal mechanism of the combination of Cinnamomum verum and Pelargonium graveolens essential oils with fluconazole against pathogenic Candida strains. Appl Microbiol Biotechnol. 2017; 101(18):6993—7006. https://doi.org/10.1007/s00253-017-8442-y
  22. Khan MS, Malik A, Ahmad I. Anti-candidal activity of essential oils alone and in combination with amphotericin B or fluconazole against multi-drug resistant isolates of Candida albicans. Med Mycol. 2012; 50(1):33—42. https://doi.org/10.3109/13693786.2011.582890
  23. Pandurang MS, Devrao HS, Ganpatrao BR, Mohan KS. Lemongrass oil components synergistically activates fluconazole against biofilm forms of Candida albicans. J Bacteriol Mycol. 2018; 5(3):1069.
  24. Miranda-Cadena K, Marcos-Arias C, Perez-Rodriguez A, Cabello-Beitia I, Mateo E, Sevillano E, et al. In vitro and in vivo anti-Candida activity of citral in combination with fluconazole. J Oral Microbiol. 2022; 14(1):2045813. https://doi.org/10.1080/20002297.2022.2045813