Abstract
The activity of six essential oils was investigated against eight fungal isolates (four Aspergillus and four Penicillium species) isolated from cultural heritage conservation premises in Serbia. To analyze the chemical composition of essential oils (EOs), gas chromatography coupled with mass spectrometry was employed. The antifungal activity of selected EOs was investigated using microdilution and microatmosphere methods while the commercial biocide benzalkonium chloride (BAC) was used as a control. Furthermore, molecular docking was used as an efficacious in silico method for the determination of interaction between dominant EO compounds and enzyme CYP51, essential for fungal ergosterol synthesis. It was demonstrated that BAC, Cinnamomum zeylanicum, and Gaultheria procumbens EOs had the strongest antifungal activity, which is in concordance with the results of molecular docking. Namely, the highest energy of enzyme–cofactor interaction was obtained for eugenol (the dominant component of Syzygium aromaticum and C. zeylanicum EOs). Moreover, it was found that the most resistant fungal isolates were A. flavus and A. niger, while A. sydowii and P. citrinum were the most susceptible. The results of our study point to the possibility of using studied environmentally friendly biocides of biological origin for the preservation of historical monuments and artifacts.


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References
Adams RP (2007) Identification of essential oil components by gas chromatography/mass spectrometry, 4th edn. Allured Publishing Corporation, Carol Stream
Anulika NP, Ignatius EO, Raymond ES, Osasere OI, Abiola AH (2016) The chemistry of natural product: plant secondary metabolites. Int J Technol Enhanc Emerg Eng Res 4(8):1–9
Barresi G, Carlo E, Trapani MR, Parisi MG, Chillè C, Mulè MF, Cammarata M, Palla F (2015) Marine organisms as source of bioactive molecules applied in restoration projects. Heritage Sci 3:17–20
Barresi G, Cammarata M, Palla F (2017) Biocide. In: Palla F, Barresi G (eds) Biotechnology and conservation of cultural heritage. Springer, Berlin, pp 49–65
Becher R, Wirsel SG (2012) Fungal cytochrome P450 sterol 14α-demethylase (CYP51) and azole resistance in plant and human pathogens. Appl Microbiol Biotechnol 95:825–840
Borrego S, Valdés O, Vivar I, Lavin P, Guiamet P, Battistoni P, Gómez de Saravia S, Borges P (2012) Essential oils of plants as biocides against microorganisms isolated from Cuban and Argentine Documentary heritage. https://doi.org/10.5402/2012/826786
Burt S (2004) Essential oils: their antibacterial properties and potential applications in foods—a review. Int J Food Microbiol 94:223–253
Candan F, Unlu M, Tepe B, Daferera D, Polissiou M, Sökmen A, Akpulat HA (2003) Antioxidant and antimicrobial activity of the essential oil and methanol extracts of Achillea millefolium subsp. millefolium Afan. (Asteraceae). J Ethnopharmacol 87:215–220
Caneva C, Nugari MP, Salvadori O (2008) Chemical methods. In: Caneva G, Nugari MP, Nugari MP, Salvadori O (eds), Plant biology for cultural heritage: biodeterioration and conservation, Getty Conservation Institute, Los Angeles, pp 99–100
Casiglia S, Bruno M, Scandolera E, Senatore F, Senatore F (2015) Influence of harvesting time on composition of the essential oil of Thymus capitatus (L.) Hoffmanns. & Link. growing wild in northern Sicily and its activity on microorganisms affecting historical art crafts. Arabian J Chem 12(8):2704–2712
Che X, Sheng C, Wang W, Cao Y, Xu Y, Ji H, Dong G, Miao Z, Yao J, Zhang W (2009) New azoles with potent antifungal activity: design, synthesis and molecular docking. Eur J Med Chem 44:4218–4226
Chinou I (2008) Primary and secondary metabolites and their biological activity. In: Waksmundzska-Hajnos M, Sherma J, Kowalska T (eds) Thin layer chromatography in phytochemistry. CRC Press, Taylor and Francis Group, Boca Raton, pp 59–76
Christoph F, Kaulfers PM, Stahl-Biskup E (2000) A comparative study of the in vitro antimicrobial activity of tea tree oils s.l. with special reference to the activity of β-triketones. Planta Med 66:556–560
Chung YJ, Lee KS, Han SH, Kang D III, Lee MH (2001) The use of fungicide and insecticide from medicinal plants for the conservation of cultural property. Conserv Stud 22:5–26
Cirone M, Figoli A, Galiano F, La Russa MF, Macchia A, Mancuso R, Ricca M, Rovella N, Taverniti M, Ruffolo SA (2023) Innovative methodologies for the conservation of cultural heritage against biodeterioration: review. Coatings 13(12):1986
Císarová M, Tančinová D, Medo J, Kačániová M (2016) The in vitro effect of selected essential oils on the growth and mycotoxin production of Aspergillus species. J Environ Sci Health, Part B 51:668–674
Cooke M (2002) European review of biocides. PharmaChem 1(6):48–50
Cuba R (2001) Toxicity myths essential oils and their carcinogenic potential. Int J Aromather 11(2):76–83
de Billerbeck VG, Roques CG, Bessière JM, Fonvieille JL, Dargent R (2001) Effects of Cymbopogon nardus (L.) W. Watson essential oil on the growth and morphogenesis of Aspergillus niger. Can J Microbiol 47:9–17
de la Paz J, Larionova M, Maceira MA, Borrego SF, Echevarría E (2006) Control of biodeterioration using a fraction isolated from leaves of Ricinus communis Linn. Pharmacol Online 3:462–466
de Leo F, Isola D (2022) The role of fungi in biodeterioration of cultural heritage: new insights for their control. Appl Sci 12(20):10490
de Martino L, de Feo V, Fratianni F, Nazzaro F (2009) Chemistry, antioxidant, antibacterial and antifungal activities of volatile oils and their components. Nat Prod Commun 4:1741–1745
Douglas MH, van Klink JW, Smallfield BM, Perry NB, Anderson RE, Johnstone P, Weavers RT (2004) Essential oils from New Zealand manuka: triketone and other chemotypes of Leptospermum scoparium. Phytochemistry 65:1255–1264
Dubey NK, Tripathi P, Singh HB (2000) Prospects of some essential oils as antifungal agents. J Med Aromat Plant Sci 22:350–354
Dupont S, Lemetais G, Ferreira T, Cayot P, Gervais P, Beney L (2012) Ergosterol biosynthesis: a fungal pathway for life on land? Evol Int J Org Evol 66:2961–2968
Džamić A, Soković M, Ristić MS, Grijić-Jovanović S, Vukojević J, Marin PD (2009) Chemical composition and antifungal activity of Illicium verum and Eugenia caryophyllata essential oils. Chem Nat Compd 45:259–261
Fierascu I, Ion RM, Radu M, Dima SO, Bunghez IR, Avramescu SM, Fierascu RC (2014) Comparative study of antifungal effect of natural extracts and essential oils of Ocimum basilicum on selected artefacts. Rev Roum Chim 59:207–211
Gadd GM, Fomina M, Pinzari F (2024) Fungal biodeterioration and preservation of cultural heritage, artwork, and historical artifacts: extremophily and adaptation. Microbiol Mol Biol Rev 88(1):e00200-e222
Ghalem BR, Mohamed B (2008) Antibacterial activity of leaf essential oils of Eucalyptus globulus and Eucalyptus camaldulensis. Afr J Pharm Pharmacol 2:211–215
Ghosh S (2015) Microwave spectroscopic study of methyl salicylate and water clusters. Master’s thesis, University of Alberta, Department of Chemistry
Gende LB, Floris I, Fritz R, Eguaras MJ (2008) Antimicrobial activity of cinnamon (Cinnamomum zeylanicum) essential oil and its main components against Paenibacillus larvae from Argentine. Bull Insectol 61:1–4
Gómez de Saravia SG, de la Paz NJ, Guiamet P, Arenas P, Borrego SF (2008) Biocide activity of natural extracts against microorganisms affecting archives. Bol Latinoam Caribe Plants Med Aromat 7:25–29
Gomez-Villalba LS, Salcines C, Fort R (2023) Application of inorganic nanomaterials in cultural heritage conservation, risk of toxicity, and preventive measures. Nanomaterials 13(9):1454
Guiamet PS, Gómez de Saravia SG (2005) Laboratory studies of biocorrosion control using traditional and environmentally friendly biocides: an overview. Lat Am Appl Res 35:295–300
Hanel H, Raether W (1988) A more sophisticated method of determining the fungicidal effect of water-insoluble preparations with a cell harvester, using miconazole as an example. Mycoses 31:148–154
Halperin I, Ma B, Wolfson H, Nussinov R (2002) Principles of docking: an overview of search algorithms and a guide to scoring functions. Proteins: Struct Funct Bioinf 47:409–443
Hameed IH, Altameme HJ, Mohammed GJ (2016) Evaluation of antifungal and antibacterial activity and analysis of bioactive phytochemical compounds of Cinnamomum zeylanicum (cinnamon bark) using gas chromatography-mass spectrometry. Orient J Chem 32:1769–1788
Hargrove TY, Kim K, Soeiro MDNC, da Silva CF, Batista DDGJ, Batista MM, Yazlovitskaya EM, Waterman MR, Sulikowski GA, Lepesheva GI (2012) CYP51 structures and structure-based development of novel, pathogen-specific inhibitory scaffolds. Int J Parasitol: Drugs Drug Resist 2:178–186
Inouye S, Watanabe M, Nishiyama Y, Takeo K, Akao M, Yamaguchi H (1998) Antisporulating and respiration-inhibitory effects of essential oils on filamentous fungi. Mycoses 41:403–410
Ishii H (1995) Monitoring of fungicide resistance in fungi: biological to biochemical approaches. In: Simgh SU, Singh PR (eds) Molecular methods in plant pathology. CRC Press, Lewis Publishers, Boca Raton, London, Tokyo, pp 483–495
James DG, Price TS (2004) Field-testing of methyl salicylate for recruitment and retention of beneficial insects in grapes and hops. J Chem Ecol 30:1613–1628
Karaalp C, Yurtman AN, Karabay Yavasoglu NU (2009) Evaluation of antimicrobial properties of Achillea L. flower head extracts. Pharm Biol 47:86–91
Kokoska L, Polesny Z, Rada V, Nepovim A, Vanek T (2002) Screening of some Siberian medicinal plants for antimicrobial activity. J Ethnopharmacol 82:51–53
Kitchen DB, Decornez H, Furr JR, Bajorath J (2004) Docking and scoring in virtual screening for drug discovery: methods and applications. Nat Rev Drug Discov 3:935–949
Koestler RJ, Santoro ED (1990) Biodeterioration in museums—observations. In: Llewellyn GC, O’Rear CE (eds) Biodeterioration Research 3. Springer science + Business media LLC, New York, pp 505–509
Kumar R, Kumar AV (1999) Biodeterioration of stone in tropical environments: an overview. The Getty Conservation Institute, Los Angeles
Langsrud S, Sundheim G, Borgmann-Strahsen R (2003) Intrinsic and acquired resistance to quaternary ammonium compounds in food-related Pseudomonas spp. J Appl Microbiol 95:874–882
Lavin P, de Saravia SG, Guiamet P (2016) Scopulariopsis sp. and Fusarium sp. in the documentary heritage: evaluation of their biodeterioration ability and antifungal effect of two essential oils. Microb Ecol 71:628–633
Lepesheva GI, Waterman MR (2007) Sterol 14α-demethylase cytochrome P450 (CYP51), a P450 in all biological kingdoms. Biochim Biophys Acta Gen Subj 1770:467–477
Ljaljević Grbić M, Unković N, Dimkić I, Janaćković P, Gavrilović M, Stanojević O, Stupar M, Lj V, Jelikić A, Stanković S, Vukojević J (2018) Frankincense and myrrh essential oils and burn incense fume against micro-inhabitants of sacral ambients. Wisdom of the ancients? J Ethnopharmacol 219:1–14
Maruzzella JC, Sicurella NA (1960) Antibacterial activity of essential oil vapors. J Am Pharm Assoc 49:692–694
Merchel Piovesan Pereira B, Tagkopoulos I (2019) Benzalkonium chlorides: uses, regulatory status, and microbial resistance. Appl Environ Microbiol 85(13):e00377-e419
Mullapudi S, Siletzky RM, Kathariou S (2008) Heavy-metal and benzalkonium chloride resistance of Listeria monocytogenes isolates from the environment of turkey-processing plants. Appl Environ Microbiol 74:1464–1468
Nagashima A, Nagato T, Kobori T, Nagi M, Okochi Y (2023) Uncommon occurrence of pulmonary Aspergillosis caused by Aspergillus sydowii: a case report. Cureus 15(12):e51353
Orlita A (2004) Microbial biodeterioration of leather and its control: a review. Int Biodeterior Biodegrad 53:157–163
Park MJ, Gwak KS, Yang I, Choi WS, Jo HJ, Chang JW, Jeung EB, Choi IG (2007) Antifungal activities of the essential oils in Syzygium aromaticum (L.) Merr. et Perry and Leptospermum petersonii Bailey and their constituents against various dermatophytes. J Microbiol 45(5):460–465
Papandreou V, Magiatis P, Chinou I, Kalpoutzakis E, Skaltsounis AL, Tsarbopoulos A (2002) Volatiles with antimicrobial activity from the roots of Greek Paeonia taxa. J Ethnopharmacol 81:101–104
Pawar VC, Thaker VS (2006) In vitro efficacy of 75 essential oils against Aspergillus niger. Mycoses 49(4):316–323
Pinto E, Vale-Silva L, Cavaleiro C, Salgueiro L (2009) Antifungal activity of the clove essential oil from Syzygium aromaticum on Candida, Aspergillus and dermatophyte species. J Med Microbiol 58:1454–1462
Porter NG, Wilkins AL (1999) Chemical, physical and antimicrobial properties of essential oils of Leptospermum scoparium and Kunzea ericoides. Phytochemistry 50:407–415
Rath CC, Devi S, Dash SK, Mishra RK (2008) Antibacterial potential assessment of Jasmine essential oil against E. coli. Indian J Pharm Sci 70:238–241
Rotolo V, Barresi G, Di Carlo E, Giordano A, Lombardo G, Crimi E, Costa E, Bruno M, Palla F (2016) Plant extracts as green potential strategies to control the biodeterioration of cultural heritage. Int J Conserv Sci 2:839–846
Salem MZ, Zidan YE, Mansour MM, El Hadidi NM, Elgat WAA (2016) Antifungal activities of two essential oils used in the treatment of three commercial woods deteriorated by five common mold fungi. Int Biodeterior Biodegrad 106:88–96
Savković ŽD, Stupar MČ, Ljaljević Grbić MV, Vukojević JB (2016) Comparison of anti-Aspergillus activity of Origanum vulgare L. essential oil and commercial biocide based on silver ions and hydrogen peroxide. Acta Bot Croat 75:121–128
Savković Ž, Stupar M, Unković N, Ivanović Ž, Blagojević J, Vukojević J, Ljaljević Grbić M (2019) In vitro biodegradation potential of airborne Aspergilli and Penicillia. Sci Nat 106:8
Sharma R, Gassel S, Steiger S, Xia X, Bauer R, Sandmann G, Thines M (2015) The genome of the basal agaricomycete Xanthophyllomyces dendrorhous provides insights into the organization of its acetyl-CoA derived pathways and the evolution of Agaricomycotina. BMC Genomics 16:233
Shulaev V, Silverman P, Raskin I (1997) Airborne signalling by methyl salicylate in plant pathogen resistance. Nature 385:718–721
Simić A, Soković MD, Ristić M, Grujić-Jovanović S, Vukojević J, Marin PD (2004) The chemical composition of some Lauraceae essential oils and their antifungal activities. Phytother Res 18:713–717
Simić A, Rančic A, Soković MD, Ristić M, Grujić-Jovanović S, Vukojević J, Marin PD (2008) Essential oil composition of Cymbopogon winterianus and Carum carvi and their antimicrobial activities. Pharm Biol 46:437–441
Shahi SK, Patra M (2003) Microbially synthesized bioactive nanoparticles and their formulation active against human pathogenic fungi. Rev Adv Mater Sci 5:501–509
Souza NAB, Lima EDO, Guedes DN, Pereira FDO, Souza ELD, Sousa FBD (2010) Efficacy of Origanum essential oils for inhibition of potentially pathogenic fungi. Braz J Pharm Sci 46:499–508
Spruijtenburg B, Rezusta A, Houbraken J, Hagen F, de Groot T, Meis JF, Meijer EF (2024) Susceptibility testing of environmental and clinical Aspergillus sydowii demonstrates potent activity of various antifungals. Mycopathologia 189(4):61
Sterflinger K, Pinzari F (2012) The revenge of time: fungal deterioration of cultural heritage with particular reference to books, paper and parchment. Minireview Environ Microbiol 14:559–566
Stojanović G, Radulović N, Hashimoto T, Palić R (2005) In vitro antimicrobial activity of extracts of four Achillea species: the composition of Achillea clavennae L. (Asteraceae) extract. J Ethnopharmacol 101:185–190
Stupar M, Ljaljević Grbić M, Džamić A, Unković N, Ristić M, Jelikić A, Vukojević J (2014) Antifungal activity of selected essential oils and biocide benzalkonium chloride against the fungi isolated from cultural heritage objects. S Afr J Bot 93:118–124
Tajik H, Jalali FSS, Sobhani A, Shahbazi Y, Zadeh MS (2008) In vitro assessment of antimicrobial efficacy of alcoholic extract of Achillea millefolium in comparison with penicillin derivatives. J Anim Vet Adv 7:508–511
Thanaboripat D, Suvathi Y, Srilohasin P, Sripakdee S, Patthanawanitchai O, Charoensettasilp S (2007) Inhibitory effect of essential oils on the growth of Aspergillus flavus. Curr Appl Sci Technol 7(1):1–7
To MS, Favrin S, Romanova N, Griffiths MW (2002) Post adaptational resistance to benzalkonium chloride and subsequent physicochemical modifications of Listeria monocytogenes. Appl Environ Microbiol 68:5258–5264
Tsai HF, Wheeler MH, Chang YC, Kwon-Chung KJ (1999) A developmentally regulated gene cluster involved in conidial pigment biosynthesis in Aspergillus fumigatus. J Bacteriol 181:6469–6477
Tyagi P, Verma RK, Jain N (2021) Fungal degradation of cultural heritage monuments and management options. Curr Sci 121(12):1553–1560
Unlu M, Ergene E, Unlu GV, Zeytinoglu HS, Vural N (2010) Composition, antimicrobial activity and in vitro cytotoxicity of essential oil from Cinnamomum zeylanicum Blume (Lauraceae). Food Chem Toxicol 48(11):3274–3280
Van Der Linden JW, Warris A, Verweij PE (2011) Aspergillus species intrinsically resistant to antifungal agents. Med Mycol 49(Suppl 1):S82–S89
Velluti A, Sanchis V, Ramos AJ, Egido J, Marín S (2003) Inhibitory effect of cinnamon, clove, lemongrass, oregano and palmarose essential oils on growth and fumonisin B1 production by Fusarium proliferatum in maize grain. Int J Food Microbiol 89:145–154
Vengurlekar S, Sharma R, Trivedi P (2014) Synthesis, antifungal activity and molecular docking studies on N-(substituted-benzylidineamine)-3-cycloalkylidine-thiosemicarbazide derivatives. Int J Drug Delivery 6:99–112
Wheeler MH, Bell AA (1988) Melanins and their importance in pathogenic fungi. In: McGiniss MR (ed) Current topics in medical mycology. Springer-Verlag, New York, pp 338–387
Wojtunik-Kulesza KA (2022) Toxicity of selected monoterpenes and essential oils rich in these compounds. Molecules 27(5):1716
Wu J, Ni T, Chai X, Wang T, Wang H, Chen J, Jin Y, Zhang D, Yu S, Jiang Y (2018) Molecular docking, design, synthesis and antifungal activity study of novel triazole derivatives. Eur J Med Chem 143:1840–1846
Xu Y, He Y, Li X, Gao C, Zhou L, Sun S, Pang G (2013) Antifungal effect of ophthalmic preservatives phenylmercuric nitrate and benzalkonium chloride on ocular pathogenic filamentous fungi. Diagn Microbiol Infect Dis 75:64–67
Young ME, Alakomi HL, Fortune L, Gorbushina AA, Krumbein WE, Maxwell I, McCullagh C, Robertson P, Saarela M, Valero J, Vendrell M (2008) Development of a biocidal treatment regime to inhibit biological growths on cultural heritage: BIODAM. Environ Geol 56:631–641
Zotti M, Agnoletti AF, Vizzini A, Cozzani E, Parodi A (2015) Onychomycosis from Aspergillus melleus, a novel pathogen for humans. Fungal identification and in vitro drug susceptibility. Exp Dermatol 24(12):966–968
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This research was supported by the Ministry of Science, Technology and Innovation of the Republic of Serbia; Grant No. 451–03 - 47/2023–01/200178.
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Savković, Ž., Džamić, A., Veselinović, J. et al. Exploring the potential of essential oils against airborne fungi from cultural heritage conservation premises. Sci Nat 112, 32 (2025). https://doi.org/10.1007/s00114-025-01983-3
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DOI: https://doi.org/10.1007/s00114-025-01983-3