Provider: - Institution: - Data provided by Europeana Collections- The application of medicinal plants and their preparations for the prevention and treatment of various disorders in humans may be limited by the possible contamination with phytopathogenic fungi and mycotoxins. Risk of using chemical fungicides for the operator, the consumer and the environment, have led to increasing interest in the introduction of alternative measures in plant protection. Lately, preparations of natural origin, so-called biological control agents are in the focus of investigation. Biological control involves the use of beneficial microorganisms (bacteria, yeasts, fungi) or the products of their metabolism, as well as the application of plant extracts and essential oils in plant protection. Examining over 40 stored dried medicinal plant species the lowest microbial quality were determined for next herbal drugs: Maydis stigmata (corn silk), Mentha leaf and herb (mint herb and leaf), Urtica leaf (nettle leaf), Equisetum herb (horsetail herb) and Calendula flower (marigold flower). Although mixed infections was recorded with different types of fungus the Fusarium was noted as the most dominant genera for most tested drugs, followed by Aspergillus and Alternaria. Twelve species of the genus Fusarium was identified. In addition, species from the following genera were identified: Phoma, Cephalosporium, Nigrospora, Cladosporium, Epicoccum, Gliocladium, Myrothecium, Cercospora, Phomopsis, Verticillum, Dreschlera (=Bipolaris), Rhizoctonia, Septoria, Trichoderma, Curvularia, Stahybotrys, Trichotecium, Puccinia, Botrytis, Mucor and Rhizopus sp. depending on plant species. In order to find an effective biological control agent, we investigated the possibility of applying the essential oils and isolates of Bacillus sp. in the control of selected identified fungi. In this regard, we chose the following fungal species: Fusarium solani, F. equiseti, F. oxysporum (isolated from corn silk and marigold flower), F. tricinctum, F. semitectum, F. sporotrichioides, F. subglutinans, Aspergillus flavus, A. niger, Alternaria alternata, Penicillium sp., Chaetomium sp., Curvularia lunata, Trichoderma viride, Trichotecium roseum, Gliocladium roseum, Myrotechium verrucaria, Phoma sp., Phomopsis sp. and Verticillium dahliae. The antifungal activity and the qualitative and quantitative content of 22 essential oils were investigated (savory, oregano, thyme, rose, anise, tea tree, geranium, lavender, coriander, bergamot, violet, rosemary, pepper, cinnamon, basil, dalmatian and spanish sage, eucalyptus, lemon, orange, chamomile and vetiver). Microdilution method on microtitration plates was used for monitoring the antifungal activity of essential oils and for determination of their minimum inhibitory and fungicide concentration (MIC and MFC) in vitro. All tested essential oils showed antifungal activity on pathogenic fungi isolated from herbal drugs, but with varying degrees of inhibitory activity. Essential oils of savory, oregano and thyme, with phenols as the dominant component, were the most effective, at the lowest concentration, while chamomile and vetiver oils, and those from Citrus species, with dominant monoterpen hydrocarbons, were the least effective against pathogenic fungi. Other oils had expressed from very good to moderate antifungal activity. Among all Fusarium species, F. subglutinans, F. semitectum and F. sporotrichioides, and T. viride, A. alternata, A. flavus i Chaetomium sp. among other genera, showed very good resistance to all oils investigated. F. oxysporum (from corn silk), M. verrucaria and T. roseum were the most susceptible pathogenic fungi. According to the results of tested synergistic activity of selected essential oils, combination of essential oils exhibited better antifungal activity than when they were tested individually. Using in situ tests, we showed that the highest efficacy in reducing the total number of fungi in samples of herbal drugs exhibited essential oils of thyme, oregano and rose. Using in vitro, dual cultivation assay, we have investigated antagonistic activity of 14 Bacillus sp. isolates, against selected fungal species. Tested isolates showed various degree of inhibition against different types of plant pathogenic fungi. The best antagonism against selected phytopathogenic fungi showed isolates SS-12.6, SS-13-1, as well as SS-27.7, SS-38.3, SS-38.4 and SS-40.2. Bacillus sp. isolates SS-39.1 and SS- 39.3 manifested the weakest antagonistic effect against almost all fungi tested. Minor antagonism against most fungi exhibited Bacillus sp. isolates SS-10.7 and SS-21.7. The . vi rest of isolates (SS-6.2, SS-35.4, SS-38.2 and SS-40.6) exhibited different antifungal potential depending on the test fungi. According to the comparative analysis of the obtained results it can be said that Fusarium species were more resistant to Bacillus sp. isolates from other genera of fungi. Evaluation of lipopeptide extracts isolated from Bacillus sp. SS-12.6 and SS- 13.1 isolates showed that the lipopeptide extract from SS-12.6 was very effective in inhibition of the growth of most fungi tested, while the lipopeptide extract from isolate SS-13.1 did not exhibit a significant antagonism against the majority of tested fungi, except Penicillium sp. Examining the potential synergism between Bacillus sp. isolates SS-12.6 and SS-13.1 and the essential oils of thyme, savory and roses, against the Fusarium species, showed that the isolates were more effective in the presence of essential oils, than when tested individually, with particular exceptions. Based on the results obtained in this work we conclude that the essential oils of savory, oregano, thyme and roses, as well as Bacillus sp. isolate SS-12.6 and its lipopeptide extract, represent a good basis for the formulation of products with potential efficacy in the control of plant pathogenic fungi, either in preventing the contamination in the field, or via indoors decontamination.- Primena lekovitog bilja i njihovih preparata u prevenciji i lečenju različitih poremećaja u ljudskom organizmu može biti ograničena njihovom mogućom kontaminacijom fitopatogenim gljivama i mikotoksinima. Saznanja o riziku pri primeni hemijskih fungicida po rukovaoca, potrošača i životnu sredinu, dovela su do povećanja interesa za uvođenje alternativnih mera u zaštiti bilja, gde posebno mesto pripada preparatima prirodnog porekla tzv. agensima biološke kontrole. Biološka kontrola podrazumeva primenu korisnih mikroorganizama (bakterija, kvasaca, gljiva) ili produkata njihovog metabolizma, kao i primenu biljnih ekstrakata i etarskih ulja u zaštiti biljaka. Ispitivanjem preko 40 vrsta lekovitog bilja najlošiji mikrobiološki kvalitet utvrđen je za sledeće droge: kukuruznu svilu, list i herbu nane, list koprive, herbu rastavića i cvet nane. Iako su na svim biljnim drogama utvrđene mešovite infekcije gljivama iz različitih rodova, većina izolovanih vrsta gljiva pripada rodu Fusarium, a potom Aspergillus i Alternaria. Osim pomenutih, identifikovani su i predstavnici rodova: Penicillium, Phoma, Cephalosporium, Nigrospora, Cladosporium, Epicoccum, Gliocladium, Myrothecium, Cercospora, Phomopsis, Verticillum, Dreschlera (=Bipolaris), Rhizoctonia, Septoria, Trichoderma, Curvularia, Stahybotrys, Trichotecium, Puccinia, Botrytis, Mucor i Rhizopus sp., u zavisnosti od biljne droge. U cilju pronalaženja efikasnog biokontrolog agensa ispitivali smo mogućnost primene etarskih ulja i izolata Bacillus sp. u kontroli odabranih identifikovanih gljiva. U tom smislu, odabrali smo sledeće vrste gljiva: Fusarium solani, F. equiseti, F. oxysporum (izolovani sa kukuruzne svile i nevena), F. tricinctum, F. semitectum, F. sporotrichioides, F. subglutinans, Aspergillus flavus, A. niger, Alternaria alternata, Penicillium sp., Chaetomium sp., Curvularia lunata, Trichoderma viride, Trichotecium roseum, Gliocladium roseum, Myrotechium verrucaria, Phoma sp., Phomopsis sp. i Verticillium dahliae. Ispitivan je kvalitativni i kvantitativni sasatav, kao i antifungalna aktivnost 22 etarska ulja (čubra, origana, timijana, ruže, anisa, čajnog drveta, zdravca, lavande, korijandera, bergamota, ljubičice, ruzmarina, bibera, cimeta, bosiljka, dalmatinske i španske žalfije, eukaliptusa, limuna, narandže, kamilice i vetivera). Antifungalna aktivnost etarskih ulja određivana je u in vitro testu, mikrodilucionom metodom na mikrotitracionim pločama. Određivane su minimalne inhibitorne i minimalne fungicidne koncentracije (MIC i MFC). Sva testirana etarska ulja su pokazala antifungalnu aktivnost na sve fitopatogene gljive izolovane sa biljnih droga, ali sa različitim stepenom inhibitorne aktivnosti. Etarska ulja čubra, origana i timijana, sa fenolima kao dominantnim komponentama, pokazala su najveći antifungalni potencijal na sve testirane gljive, pri najnižim koncentracijama. Etarska ulja vetivera i kamilice, sa seskviterpenima kao dominantnim komponentama, su pokazala najslabiju antifungalnu aktivnost, kao i etarska ulja eukaliptusa, limuna i narandže u kojima preovladavaju monoterpenski ugljovodonici. Ostala ulja ispoljila su od veoma dobre do umerene antifungalne aktivnosti. Među vrstama roda Fusarium, F. subglutinans, F. semitectum i F. sporotrichioides, generalno su najotpornije gljive na većinu analiziranih etarskih ulja, a od predstavnika drugih rodova to su T. viride, A. alternata, A. flavus i Chaetomium sp. Najosetljivije gljive na većinu etarskih ulja su F. oxysporum izolovan sa kukuruzne svile, kao i M. verrucaria i T. roseum. Prema rezultatima ispitivanja sinergističke aktivnosti odabranih etarskih ulja može se reći da su kombinacije etarskih ulja ispoljile bolju antifungalnu aktivnost nego kada su ulja ispitivana pojedinačno. U in situ ispitivanjima, najveću efikasnost u redukciji ukupnog broja gljiva u uzorcima biljne droge, ispoljila su ulja timijana, origana i ruže. Primenom in vitro testa dvojne kultivacije ispitivana je antagonistička aktivno