Ethnopharmacology is the study of folk (local) and traditional use of plants, animals, fungi, microorganisms and minerals for prevention and treatment of diseases in humans and animals. Ethnopharmacological research usually begins with a field research in a local community, in which researchers document the use of folk (local) or traditional medicines. Field research is followed by a phytochemical, pharmacological and toxicological evaluation of medicines with the aim of contributing to their better and safer use. Karst and Gorjanci (Dolenjska) are two predominantly rural Slovenian areas that are approximately 120 km apart. Karst is located near the Italian border and Gorjanci near the Croatian border. Karst has a climate with Mediterranean and continental influences, and Gorjanci has a moderate continental climate. In the past, the inhabitants of both regions were farmers. Today, however, the active population works in the nearby industrial centres. Nevertheless, people are still connected to nature and knowledge about plants is important in their lives because they produce their own food in gardens or fields and some are still farmers. 25 informants4 were interviewed between October 2013 and September 2014 in the ethnopharmacological field research in Karst and Gorjanci, respectively. The main question was: "Which plants do or did you collect from nature and use?5" Plants for medicinal, nutritive, veterinary or cosmetic use were considered. Informants reported 77 different plants in Karst and 81 in Gorjanci; 64 plants were reported in both regions. Approximately 64% of the plants in each region were distinctive for only a few informants (fewer than 7). The remaining plants were considered important, and the majority was mutual to both regions. Few reported plants were typical for just one region. These differences were mostly due to the ecological distribution of wild plants and suitable growing conditions of cultivated plants, which differed between both regions. The most frequently reported uses of plants were medicinal and nutritive, and between both uses, a continuous passage was present. For aproximately 50% of plants, medicinal and nutritive uses were reported. The informants also frequently mentioned properties that promote health, e.g. a high content of vitamin C, although the plants were not (necessarily) used for medicinal purposes. The highest number of plants was used for the treatment of gastrointestinal disorders, respiratory system disorders and skin diseases. Differences in the reported medicinal purpose of use for some frequently collected plants were observed between Karst and Gorjanci. These regional differences could be explained by cultural differences, which were possibly due to to the influences from the neighboring areas. Oral sources were also important providers of botanical knowledge to the informants. The informants were mostly elderly people with a mean age of 61 years in Karst and 69 years in Gorjanci. However, their knowledge about plants seemed to be influenced by media, most often popular books about medicinal plants that were published in the 20th century. Traditional use of fly agaric (Amanita muscaria) for catching flies was also investigated in Karst and Gorjanci. Fly agaric has been used for this purpose for centuries. Nevertheless, such preparations are poorly documented and researched. To the best of our knowledge, the oldest record of the use of fly agaric for catching flies is in the book Flora carniolica (1760) written by Antonio Scopoli. In Karst, only three informants reported the use of fly agaric for catching flies and provided a detailed description of the preparations used. These three informants were originally from other parts of Slovenia and remembered the use from their home. In Gorjanci, 13 informants provided a detailed description of fly agaric preparations. Results suggest that the tradition of preparing fly agaric for catching flies was present in Gorjanci but not in Karst. Traditional knowledge, which is still present, will likely disappear in the coming years due to the lack of reports about current use. Infromants reported a total of nine different methods for preparing fly agaric for catching flies. Some methods were simple and included soaking in milk or water or dripping a little milk onto the mushroom. Others were more complex and included a combination of heat or mechanical processing and soaking in milk or water. Eight preparations were prepared according to the traditional methods, and the release of ibotenic acid and muscimol from the fungal material was determined by HPLC in five different time points (0.5, 1, 2, 3 and 24 h). For all preparations, the release of ibotenic acid was time-dependent, with the extracted amount increasing over time. Although milk was used more often than water in traditional recipes, the release of both substances was not dependent on the solvent used. Fungal material that was exclusively soaked in water or milk released the smallest amount of ibotenic acid and muscimol at each time point. Additional heat and mechanical processing led to faster release of ibotenic acid and muscimol from the fungal material. In the third part of the doctoral work, we studied the use of aconite (Aconitum spp.) in Solčavsko region6. Aconitum species are poisonous plants that were part of official and folk medicine in the Slovenian territory in the 19th century. Due to its toxicity, the plant stopped being used as official medicine in the 20th century. Results of the ethnopharmacological field investigation indicate that folk knowledge of the medicinal use of Aconitum spp. is still present in Solčavsko, although the plant is used much less frequently than in the past. The informants reported the preparation of ethanolic extract made from homemade spirits and aconite roots, most likely from the species Aconitum tauricum and Aconitum napellus. The infomants did not report about the use of aconite herb. The official medicine begun to use the aconite roots in the second half of the 19th century but until than, only aconite herb was used. The present use of ethanolic extract made from aconite roots in Solčavsko indicates a possible influence of official medicine on the folk use of this plant in this area. The extract was used internally and externally for various indications; use in animals was also reported. Four informants kept the extract at home; two extracts were prepared recently (1998 and 2015). By phytochemical analysis, we confirmed the presence of aconitine in two out of three home-made extracts; the fourth was not analysed. There was no general rule for dosing of the extract, since each informant reported a different number of administered drops. Although the recipes for the extract and the reported dosages differed greatly among the informants, no poisonings due to the medicinal use of the extract were reported. In the past, the folk use of plants was in constant change because the knowledge of their use was continually being built, changed, transmitted and lost. Advancement in technology today enables us to find new knowledge about plants faster than ever before, as well as transmission of new and old knowledge about medicinal plants to a broader public. This in turn leads to even more rapid changes and possibly unification of the local use of medicinal plants. With this research, we contributed towards greater knowledge of the present folk use of plants and fungi, specifically fly agaric, in Karst and Gorjanci, and the use of aconite in Solčavsko. The findings of this research are a snapshot of the current folk knowledge about plants, which originates in our history, affects our society today and will affect us in the future.
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