Journal of Ethnopharmacology 151 (2014) 704–713

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Traditional use of the native medicinal plant resource of Mt. Rtanj (Eastern Serbia): Ethnobotanical evaluation and comparison Bojan K. Zlatković a,n, Stefan S. Bogosavljević b, Aleksandar R. Radivojević c, Mila A. Pavlović d a

Department of Biology and Ecology, Faculty of Sciences and Mathematics, Višegradska 33, University of Niš, Serbia Pharmaceutical Association of Serbia, Dečanska 8a, Belgrade, Serbia c Department of Geography, Faculty of Sciences and Mathematics, Višegradska 33, University of Niš, Serbia d Faculty of Geogrphy, Studentski Trg 3/1, Univeristy of Belgrade, Serbia b

art ic l e i nf o

a b s t r a c t

Article history: Received 8 September 2013 Received in revised form 4 November 2013 Accepted 21 November 2013 Available online 1 December 2013

Ethnopharmacological relevance: This study reports on the uses of medicinal plants and ethnobotanical significance of Mt. Rtanj, Eastern Serbia. The aim of the study was collecting information on plant utilization and creating the list of medicinal plants collected by local community in the region of Rtanj Mt. in Serbia. Methodology: A group of generally elderly people from the region were interviewed, asked to identify a certain number of plant taxa used against various human ailments. A quantitative ethnobotanical comparison has been performed in relation to other studies concerning the central Balkan Peninsula. Results: There is an inventory of 45 plant species recorded. At the family level, the most common in traditional usage were species of Labiatae (22%), Rosaceae (20%) and Compositae (13%). The results of the survey suggested that Hypericum perforatum was the most popular medicinal plant. Among the 7 different recorded types of preparation, the aerial parts of plants were the most popular (47.3%). The most frequently reported medicinal uses of herbal drugs include remedies for immune system (22.97%), respiratory system (15.77%) and digestive system disorders (15.32%). However, a comparatively small number of available plant species was used for medicinal purposes while the level of ethnobotanical diversity was lower when compared to adjacent regions. Conclusions: Ethnobotanical knowledge in this area is decreasing due to high emigration rates during the last period. The historically developed ethnobotanical heritage should be preserved and promoted on the larger level, getting the special position in the future management plans for this area. & 2013 Elsevier Ireland Ltd. All rights reserved.

Keywords: Rtanj Mt. Ethnobotany Balkan Peninsula Medicinal plant resource Comparison

1. Introduction According to the latest data, vascular flora of Serbia includes about 3662 plant taxa (species and subspecies), placing it in the group of countries with prominent richness of vascular plants in Europe (Stevanović et al., 1999). This area is part of the Balkan Peninsula, representing the concurrence region of most diverse historical and biogeographical influences (Stevanović et al., 1999; Turrill, 1929). In fact, plant life in the whole Peninsula represents the final stage of long-term evolutionary and plant migration processes that have created the original and rich flora (Turrill, 1929). Plant species used in traditional medicine and for other

n

Corresponding author. Tel.: þ 38118533015x55. E-mail addresses: [email protected] (B.K. Zlatković), [email protected] (S.S. Bogosavljević), [email protected] (A.R. Radivojević), [email protected] (M.A. Pavlović). 0378-8741/$ - see front matter & 2013 Elsevier Ireland Ltd. All rights reserved. http://dx.doi.org/10.1016/j.jep.2013.11.037

ethnobotanical purposes represent a large portion of its floristic richness. Summarized data from different studies have shown that about 700 wild growing species have been used as medicinal plants of Serbia (Sarić, 1989). Many studies have been performed in the mountain regions of the Balkan Peninsula in recent times (Pieroni et al., 2005; Pieroni, 2008; Jarić et al., 2007; Šarić-Kundalić et al., 2010, 2010a; Menković et al., 2011; Pieroni et al., 2011; Mustafa et al., 2012, 2012a; Rexhepi et al., 2013; Šavikin et al., 2013), elucidating the importance and traditional use of its wild plant resources. Rtanj Mt. belongs to the group of best known mountains among the general public regarding the richness and originality of its medicinal flora. The tradition of harvesting medicinal plants in this region is very long, representing a special segment of the tourism market demand (Jovanović and Radivojević, 2006). Collecting herbs is valorized through different tourist events, cultural and herbalist manifestations (Milenković, 2004). It is necessary to emphasize the efforts of numerous local nongovernmental,

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governmental and international organizations that have played an important role in the promotion and conservation of available natural resources and medicinal flora of Rtanj region during the last decade. The single ethnobotanical study in this region, written in form of an essay, was published more than two decades ago by Milojević (1988), providing information on some 80 species referred by their medicinal and other applications. However, information in the previous study was collected from randomly contacted informants and passers-by in the mountain, asked to choose within a heap of collected material from the field. Considering the richness and originality of Balkan flora, promotion of original selection and type of use of medicinal plants might be built-in in economic development and commercial use of available natural resources. For the communities living in mostly economically poor highland regions, promotion of traditional knowledge and practice means a way toward achieving a higher standard of living. This study was organized in order to increase the amount of knowledge and documentation on wild-growing medicinal flora of mountain regions in the Balkan Peninsula as one of the most important centers of plant diversity in Europe (Stevanović et al., 1999; Radford and Ode 2009), and also to promote the practical significance of use of this flora.

2. Background and methods 2.1. Study area The chain of Rtanj Mt. is situated in the C part of Balkan Peninsula belonging to the group of Carpatho–Balkan Mountains, spreading from Romania toward Eastern Serbia. The mountain covers approximately 296.4 km2 (43146′ N and 21153′ Е) which is more or less equal to the studied area (Fig. 1). As it is one of the highest mountains of E Serbia, this mountain has a more or less isolated position, leading to differences from the other mountains in geophysical sense. It is a composite system of relatively high peaks forming the northern edge of Sokobanja basin, as one of the largest of its type in the C Balkans (Gavrilović 1970; Jovanović, 1923). The broad mountain foothills arise at c. 300 m. above the sea level, up to the mountain ridge that forms a line about 36 km

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long, including several prominent peaks, and with relative NE–SW direction. Due to its specific landmarks Rtanj is very distinct from the other elements of the Carpatho-Balkan mountain arc. First of all, this massif has original geological and tectonic traits when compared to the other members. In geological terms this region is characterized by complex composition. Deposits of mainly sedimentary Mesozoic origin have dominant distributions. Mighty limestone rocks participate in construction of higher horizons and are mostly limited to the peaks of the ridge (Pavlović et al., 2011). The broader region around Rtanj, including the surrounding basins and valleys, is characterized by modified temperatecontinental climate, with pluviometric regime affected by Mediterranean climatic influence. The temperature of the coldest month is above  3 1C, while the mean temperature of the warmest period is generally below 22 1C (Rakićević, 1980). Due to the prominent relief the climate in the higher regions of Rtanj is moderately modified and colder. The pluviometric regime is generally conditioned by changes in altitude. While the base of the massif receives 711 mm, the highland area receives about 1459 mm of total annual precipitation. The geographical position and other natural conditions have caused the plant life of Rtanj Mountain to be one of the most interesting in the region. The dominant limestone substrate together with prominent relief has a considerable influence on composition and distribution of flora and vegetation. The northern mountain slopes are characterized by diverse and relatively well-developed forest vegetation. Wide, strongly eroded areas of southern mountain slopes are covered in dense shrub and grassy vegetation with a rich and diverse floristic composition (Jovanović-Dunjić, 1956). Vegetation of rock fissures typical for limestone-based mountains is developed in the higher part of the mountain and represented by communities quite local in distribution. According to Jovanović-Dunjić (1956), the transitional climate position of the mountain is important for the survival of sensu lato Mediterranean species inhabiting the same habitats as the abundant steppe species (Diklić 1965; Diklić and Milojević, 1976). Flora of this mountain is distinguished by the presence of relict species such as Ramonda serbica, Viola grisebachiana and Corylus colurna originating in Tertiary age, as well as by rare steppe elements

Fig. 1. Map of the study area with main mountain peaks and water bodies highlighted: () villages visited in the scope of ethnobotanical interviews.

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(Prunus tenella, Pulsatilla montana, Paeonia mascula, Adonis vernalis). They are important as issues of ecological and conservational concern, providing great scientific and practical importance to the flora of this mountain, as well as the whole Balkan Peninsula. Regarding the originality of nature, it is also important to consider this area from the conservational point of view. Being extremely important from the aspect of biodiversity conservation, the area of 87.46 ha was proclaimed to be protected according to the national legislative from 1959. Rtanj Mt. is also recognized as an important biodiversity “hot spot” at international level (Amidžić et al., 2011; Jakšić, 2008; Radford and Ode, 2009) as listed in IPA (Important Plant Areas), IBA (Important Bird Areas), PBA (Prime Butterfly Areas) and EMERALD (Areas of Special Conservational Importance-ASCI). It is worthy to note a number of species used for medicinal purposes, veterinary medicine, human and animal nourishment, as well as ritual life of the settlements in this mountain, as recorded in previous studies (Milojević, 1988). Recent studies of potentially important medicinal plants from Rtanj mountain, such as the strictly local and endangered Nepeta rtanjensis described from Rtanj Mt. (Diklić and Milojević, 1976, Diklić, 1999), reveal their chemical composition, antimicrobial and antifungal activity (Stojanović et al., 2005; Ljaljević-Grbić et al., 2008; Ljaljević-Grbić et al., 2011). However, these data should not be used to support collection of such a rare species from the natural habitats on the mountain, but instead to emphasize importance of their “in vitro” propagation in order to improve natural populations and for their later cultivation for harvesting.

2.2. Localities and population The study area is specific in all aspects of demographic development. It was extremely depopulated throughout the second half of the 20th and beginning of 21st century with a permanent decline in population numbers (Table 1). These trends have led to negative changes in all demographic structures. The age structure is particularly unfavorable and the average age of the population in the region is above 60. Natural increment is negative (  1.2‰) as in some of the studied villages the biological reproduction has almost entirely ceased (Kojić, 2007). In addition, the process of prominent migration took place since the middle of the 20th century and still exists in the area. In all the villages visited, young people under the age of twenty were a minority due to migration of the younger population to the closest larger towns, as has been a growing trend during the past decades (Radivojević et al., 2012). Unfavorable age structure affects the population structure by sectors and activities. Migration of young people from the mountain villages also indicates a potential loss of knowledge on medicinal plants and their use in the future.

According to our studies, about 18% of medicinal species recorded in previous ethnobotanical screening by Milojević (1988) were not in use any more among the community living in the same region. Decreasing trend of ethnobotanical knowledge was noticed by Jarić et al. (2007) for Kopaonik Mt. in Serbia, but it is also attributive to many other mountain regions beyond the Balkans. The population of studied villages is mainly engaged in animal husbandry and agriculture (Pavlović and Radivojević, 2009). The villages are located at different elevations (360–560 m a.s.l.). They are relatively small in terms of their given area and the number of inhabitants. The native residents are ethnic Serbs, who speak Serbian, a Slavic language.

2.3. Ethnobotanical survey Ethnobotanical survey was based on information collected during the studies conducted in the period 2011–2012. Information gathered in the form of a questionnaire provides insight into the knowledge on medicinal plants and original ways of their use among the local community at Rtanj Mt. Field surveys included the territory of Sokobanja and Boljevac districts, comprising a total number of 10 villages defined in the physical meaning of mountain settlements (Fig. 1). Expected age-related knowledge reflecting the most traditional uses was the main reason to perform sampling in the group of generally older people. Most respondents were engaged in agriculture and animal husbandry, and their knowledge of herbs and their uses was based on tradition inherited from their relatives and neighbors. Information gathering was carried out through direct interview in which each respondent was asked a list of questions. In this phase of the study, respondents were asked to specify the group of species that they collect on the mountain for medicinal purposes, and to add as much information as possible on their potential use in treatment of certain disorders. Respondents were also asked for the following information: which part of particular plant shows optimal medicinal effect, to explain the precise way for preparing herbal remedies, and finally to specify the health problem or injury for which the certain treatment type is used. The identity of reported species was checked in the field by asking the informants to show the specimen or plant parts of the exact species so they could be identified by the expert. Quotation frequency was calculated for every reported species on the base of informant statements. It refers a portion of selected species in total number of use reports (indications) reported in survey. Original notes, questionnaire data and other documented materials obtained during the field survey are deposited at Department of Geography, Faculty of Natural Sciences, University of Niš.

2.4. Plant material Table 1 Demographic trends in rural areas in the study area for the period 1953–2011. Settlement

Vrmdža Jošanica Nikolinac Ilino Mirovo Seselac Rujište Mužinac Šarbanovac Dobro Polje

Altitude (m)

460 400 500 360 400 470 560 540 570 373

Number of citizens 1953.

1961.

1971.

1981.

1991.

2002.

2011.

1568 1573 962 482 615 935 854 922 1158 1064

1479 1588 896 457 714 915 778 895 1123 998

1248 1654 798 335 512 822 628 835 994 917

1124 1595 672 216 404 664 590 762 861 756

901 1422 530 178 287 488 490 643 708 590

606 898 418 121 183 492 388 459 514 415

517 787 316 101 140 254 297 407 424 306

As reported in the questionnaire, following the propositions given by Heinrich et al. (2009), all species collected during fieldwork were labeled after identification and deposited in the herbarium collection at Department of Biology and Ecology, Faculty of Natural Sciences, University of Niš (HMN). The voucher numbers are given in the brackets after the scientific name of the taxa (Table 2). The nomenclature of the taxa listed, given at species and subspecies level, was for the most part compiled from contemporary check-lists, monographic works and databases EUROþ MED (Plantbase, http://ww2.bgbm.org/EuroPlusMed) and IOPI (http:// plantnet.rbgsyd. nsw.gov.au/iopi/iopihome.htm), but to some extent also from earlier monographs and local floras (Jordanov, 1963–1979; Josifović, 1970–1977).

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Table 2 List of medicinal plant species from Rtanj Mountain used in traditional medicine, reported from Informants. Parts used

Type of Medicinal use preparations

0.9

Herba

Infusion

Antidiabetic.

8.6

Herba

Infusion, poultice

0.9

Folium

Fresh

Appetite stimulant, sedative, against anemia, peptic ulcers, immune system strengthening, menstrual difficulties, ovarian cysts. Externally, as vasotonic, against hemorrhoids and purulent wounds. Against anemia, stimulant.

0.5

Folium

Infusion

Against cough.

Repušina, čičak Hoću-neću

0.9

Herba

Infusion

Against cough, against cold.

1.4

Herba

Infusion

As vasotonic, against hemorrhoids.

Kičica

2.3

Herba

Infusion

Antiflatulent, digestive, gastritis, antidiabetic, stimulant.

Gologuza, vodopija Đurđevak

0.5

Herba

Infusion

Digestive.

0.9

Flos

Infusion

Diuretic, antihypertensive.

Dren

3.6

Flos, fructus

Beli glog

3.6

Folium cum flores, fructus Folium cum flores, fructus Herba

Infusion, ptisan Infusion, ptisan

Against diarrhea, intestinal diseaes, against anemia, stimulant, immune system strengthening. Cardiotonic, antihypertensive, intestinal diseaes, stimulant.

Infusion, ptisan

Antihypertensive.

Infusion

Kidney diseaes, kidney stones.

Folium

Infusion

Against cough.

Folium

Infusion

Against cough.

Herba

Infusion, oil

Herba

Infusion

Immune system strengthening, peptic ulcers, stimulant. Externally, for purulent wounds, wound healing. Against cough.

Fructus

Immune system strengthening, against cold, digestive, antihypertensive.

Herba

Ptisan, vinegar Infusion

Herba

Infusion

Menstrual difficulties, against hemorrhoids.

Flos

Infusion

Folium

Infusion

Immune system strengthening, against cold, sedative, against conjuctivitis, antiinflammatory. Sedative.

Herba

Infusion

Against cold, digestive, antispasmodic.

Herba

Infusion

Menstrual difficulties, ovarian cysts.

Herba

Infusion

Immune system strengthening, digestive, hypolipemic.

Folium, radix

Infusion, poultice

Bronhitis, peptic ulcers. Externally, for purulent wounds, skin diseases, skin ulcers, wound healing.

Herba

Infusion

Stimulant.

Flos, folium

Infusion

Expectorant.

Stipites, fructus Fructus

Infusion, ptisan Ptisan

Urinary infections.

Fructus

Ptisan

Digestive, stimulant.

Fructus

Ptisan

Stimulant.

Folium, fructus

Infusion, fresh

Immune system strengthening, stimulant, against anemia, for sore throat.

Botanical taxa and family

Local name

Achillea clypeolata Sibth. and Sm. (Compositae) (6863)

Žuta hajdučka trava, mesečina Hajdučka trava

Achillea millefolium L. (Compositae) (6864) Allium ursinum L. (Liliaceae) (6865) Althaea officinalis L. (Malvaceae) (6866) Arctium lappa L. (Compositae) (6867) Capsella bursa-pastoris (L.) Medicus (Cruciferae) (6868) Centaurium erythraea Rafin. (Gentianaceae) (6869) Cichorium intybus L. (Compositae) (6870) Convallaria majalis L. (Liliaceae) (6871) Cornus mas L. (Cornaceae) (6872) Crataegus monogyna Jacq. (Rosaceae) (6873)

Crenbuš, medveđi luk Beli slez

Quotation frequency (%)

1.8 Crataegus pentagyna Waldst. & Crni glog Kit. ex Willd. (Rosaceae) (6874) 0.5 Equisetum arvense L. Rastavić, (Equisetaceae) (6875) poljski rastavić Fragaria vesca L. (Rosaceae) Divlja jagoda 0.9 (6876) Hedera helix L. (Araliaceae) Bršljan 0.5 (6877) Hypericum perforatum L. Kantarion 12.7 (Guttiferae) (6878) Hyssopus officinalis L. Vuzak, 0.5 (Labiatae) (6879) miloduh Malus sylvestris (L.) Mill. Divlja jabuka 3.6 (Rosaceae) (6880) Marrubium peregrinum L. Bela očajnica 1.4 (Labiatae) (6881) Marrubium vulgare L. Očajnica 0.9 (Labiatae) (6882) Matricaria chamomilla L. Kamilica 2.7 (Compositae) (6883) Melissa officinalis L. (Labiatae) Matočina 0.9 (6884) Mentha pulegium L. (Labiatae) Barska nana 2.3 (6885) Nepeta cataria L. (Labiatae) Macina trava 1.4 (6886) Origanum vulgare L. (Labiatae) Vranilova 1.4 (6887) trava 6.3 Plantago major L. Bokvica, (Plantaginaceae) (6888) ženska bokvica Polygonum aviculare L. Troskot 0.5 (Polygonaceae) (6889) Primula veris L. (Primulaceae) Jagorčevina, 1.4 (6890) jaglika Prunus avium L. (Rosaceae) Divlja trešnja 1.4 (6891) Crni trn, 0.9 Prunus spinosa L. (Rosaceae) (6892) trnjina Pyrus spinosa Forssk. Divlja kruška, 2.3 (Rosaceae) (6893) slanopađa Rosa canina L. (Rosaceae) Šipurak, divlja 1.8 (6894) ruža Divlja kupina 4.5 Rubus ulmifolius Schott (Rosaceae) (6897)

Menstrual difficulties, against hemorrhoids.

Against diarrhea.

708

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Table 2 (continued ) Botanical taxa and family

Local name

Rumex patientia L. Divlje zelje (Polygonaceae) (6898) Salix alba L. (Salicaceae) (6899) Divlja vrba, bela vrba Sambucus nigra L. Zova (Caprifoliaceae) (6900) Satureja kitaibelii Wierzb. ex Rtanjski čaj, Heuff. (Labiatae) (6901) čubar Sempervivum marmoreum Čuvarkuća Griseb. (Crassulaceae) (6902) Taraxacum officinale Weber Maslačak (Compositae) (6903) Teucrium montanum L. Iva, trava-iva (Labiatae) (6904) Majkina Thymus praecox Opiz subsp. dušica jankae (Čelak) Jalas (Labiatae) (6906) Tilia cordata Miller (Tiliaceae) Bela lipa (6908) Tilia platyphyllos Scop. Crna lipa (Tiliaceae) (6909) Kopriva Urtica dioica L. (Urticaceae) (6910) Veronica officinalis L. Čestoslavica (Scrophulariaceae) (6911)

Parts used

Type of Medicinal use preparations

0.5

Folium

Infusion

Against anemia.

1.4

Folium

Infusion

Bile disease.

3.2

Flos, fructus

Diaphoretic, bronhitis.

4.1

Herba

Infusion, ptisan Infusion

Immune system strengthening, against cold, for hypotension.

0.5

Folium

Fresh

Against ear infections, antiinflammatory.

1.4

Folium, flos

Infusion

Expectorant, diuretic.

0.5

Herba

Infusion

Immune system strengthening.

2.3

Herba

Infusion

Against cold, stimulant, sedative.

5

Flos

Infusion

1.8

Flos

Infusion

Immune system strengthening, against cold, against insomnia, sedative, bile diseaes. Against cold, against insomnia.

4.1

Folium

Infusion

Against anemia, stimulant, immune system strengthening.

1.4

Herba

Infusion, poultice

Against anemia, hypolipemic. Externally, for skin diseases.

Quotation frequency (%)

2.5. Data analysis All collected data from the questionnaires were incorporated into datasets, including scientific name of the plant, part(s) used, mode of use and medicinal purpose. These records were associated with source informant data including age, gender and environmental features such as altitude and geographical position of the settlement. We tested whether the choice of species reported was influenced by any of the several factors as different variables, including geographic area, altitude, age and gender of informants. We applied a General Linear Model ANOVA entering “number of taxa” and “number of reports” per responder as dependent variables. Significance was set at p ¼0.05. We also applied multivariate correspondent techniques (Rohlf, 1988) in order to reflect on potential relationship among variables, observing the state of their agglomeration in graphs. The output of this analysis shows the coordinates of the row (informants) and column (character states) on correspondence axes displayed on the scatter plot. In order to determine ethnobotanical meaning of the investigated area we have compared the data on species richness among several mountain regions of central Balkan Peninsula. To make a more reliable ethnobotanical comparison between selected districts, we performed agglomerative hierarchical clustering (AHC). The method is applied utilizing the presence of certain medicinal species in different areas as variables (aggregation criterion: unweighted pair-group average and Euclidian distance). All multivariate analyses were carried out with statistical package STATISTICA 6.0 (STATSOFT 2001). We calculated quantitative relationship (LogS/LogA) of area surface (A) and total number of reported medicinal species (S) derived from selected ethnobotanical studies (Milojević, 1988; Pieroni et al., 2005; Jarić et al., 2007; Pieroni, 2008; Menković et al., 2011; Pieroni et al., 2011; Mustafa et al., 2012, 2012a; Rexhepi et al., 2013; Šavikin et al., 2013) in order to express and compare the level of ethnobotanical diversity per region. Informant consensus factor (Fic) is used to express ethnobotanical homogeneity. It is calculated as the ratio between the number of use-reports minus the number of medicinal plants and the number of use reports minus one

(Heinrich et al., 1998). Factor ranges from 0 to 1 and higher value indicates a high rate of agreement between the informants, a low one a low degree of agreement.

3. Results and discussion 3.1. Quantitative ethnobotanical analyses 37 informants were interviewed, with both sexes almost equally represented (19 women and 18 men) whose averaged ages ( 7standard deviation) were 64.86 75.82 years (range 55–78 years). Plant data included 45 taxa (a full list with remarks is given in Table 2) from 20 different families of vascular plants. In general, 222 statements (use-reports) were obtained for all taxa, referring to 7 different plant parts and 6 methods of preparations recorded, with total number of 39 indications (medicinal uses) from 10 groups of versatile health disorders. Total number of reports exceeded the number of taxa, as each taxon often had more than one use and the same species was also mentioned by numerous respondents. Top ten replies mentioned by key informants refer to 16 different species, covering about 71.04% of total number of citations. In all cases the species most cited was Hypericum perforatum, as the most commonly used and the highly indicated herb. Nine species, or 4.05% of the total species number, were reported only once. Key informants mentioned 20 families, 6 of which were mentioned more than 10 times, making the 76.92% of all citations. At the family level the most cited group was Labiatae, represented by 10 mainly traditionally used aromatic taxa (e.g. Mentha pulegium, Thymus praecox, Nepeta cataria and Origanum vulgare) including endemic Satureja kitaibelii well-known by its vernacular name Rtanj's tea. Labiatae are followed by Rosaceae, with 9 recorded species including popular native fruits such as hawthorn, almond-leaved pear, common apple and dog-rose. The third position belongs to Compositae (6 taxa) as one of the largest families of flowering plants in the flora of the Balkan Peninsula (Turrill, 1929), that is followed by Liliaceae (2), Tiliaceae (2) and Polygonaceae (2). The

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rest of the reported families making the majority of the list (70%) were represented by single species. Reports have exclusively pertained to medicinal applications within large spectra of different indications (Table 2). On average, 2.46 71.81 medicinal uses and indications were reported for each species. Similar range was recorded regarding the 6 possible preparation methods. On average 1.2470.43 preparations were reported per species. Variance analyses ANOVA has shown no significant effects concerning the number of taxa or the number of data sets per responder (P 40.05) depending on geographic area (F(9,27) ¼1.484; p¼ 0.207), altitude (F(2,34) ¼1.648; p ¼0.162) age (F(2,34) ¼1.524; p¼ 0.194) or informant's gender (F(1,35) ¼0.26; p ¼0.874). On average, women mentioned 7.5 75.2 taxa (range 2–19); men 9.0 7 6.8 taxa (range 1–28). The analyses of local knowledge have shown dominance of a few medicinal species which were overrepresented above all others mentioned in the survey. This fact seems to be a rule as it has also been reported by Blanco et al. (1999), Idolo et al. (2010) and Camejo-Rodrigues et al. (2003) as a result of territorial continuity in many different cases. The ethnobotanical homogeneity in this region is a clear consequence of absence of significant geographical barriers between the villages. The absence of difference between villages was caused by the more or less identical altitude in which they are situated. It is also the result of frequent communication, sharing information among the communities on the mountain. Ethnobotanical investigations often emphasize the role of responder age in ethnobotanical analyses where collected knowledge belongs to persons from older generations. Age-related rate of ethnobotanical knowledge was tested in a similar way as in the study of Idolo et al. (2010). Three key groups of responders were formed according to their age (51–60; 61–70; 71–80). As it was expected, no statistical correlations were found between age categories and type of information, first of all the number of stated species. Small and relatively equal quantity of statements reported per responder may represent the result of tradition and specific relatives-chain transport of ethnobotanical information within the

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analyzed population. On the other hand, it reflects relatively solid social relationships between small local communities. High informant consensus factor (Fic) value for this region (0.80) also confirms the above facts. It is usually used to express ethnobotanical homogeneity in the studies, representing the ratio between the number of use-reports and the number of medicinal plants reported (Heinrich et al., 1998; Akerreta et al., 2007). The results of applied multivariate techniques have shown potential correlation among at least some of the 6 variables tested, indicating the specific state of their agglomeration in analyses (Fig. 2). These agglomerates are particularly important as they reflect the specific character of ethnobotanical knowledge in the studied community. Specific information refers to the originality in species choice, part of the plant in use and treatments, but it might also stress out specific or even unusual indications from the analyzed data. As a result of correspondent analyses three groups of elements were determined. The first group indicates application of oleum or green parts and inflorescences of Guttiferae (Hypericum perforatum) in treatment of skin system disorders and wound healing, and aerial parts of Scrophulariaceae (Veronica officinalis) species in treatment of skin diseases. Another group includes the use of leaves from Plantaginaceae (Plantago major) species in the form of poultice, overlapping the medicinal use with the previous group of associated elements. As it was expected, use of Hypericum oil was shown to be one of the favorite remedies, together with leaves of Plantago major which are used in variety of skin problems and injuries. Unexpected appearance of plants from family Scrophulariaceae in the first group refers to the interesting use of Veronica officinalis aerial parts in the treatment of skin diseases. In Serbian traditional medicine it has been used as a common cure for bronchitis, asthma, gastrointestinal aliments, rheumatism and arthritis, while in the past it was also used to treat skin ulcers and disorders (Sarić, 1989). However, there are also a few cases of similar external use of this species in the traditional medicine of neighboring countries. For example in Romanian folk medicine it is known for wound healing effects (Scarlat et al., 1985), and among

Fig. 2. Results of correspondent analysis indicate statistical connections among the three groups of analyzed elements, regardless of the position of settlement on the mountain, age cohort or gender of responder: (1) use of leaves (FOL) or oleum (OIL) of families Guttiferae (GUT) and Scrophulariaceae (SCR) in treatment of skin system disorders (SSD); (2) use of species from family Plantaginaceae (PLA) as poultice (POU) in treatment of skin ailments and wounds; (3) use of fresh parts (FRE) or tisane (PTI) made of fruits (FRU) from families Crassulaceae (CRA), Rosaceae (ROS) and Cornaceae (COR) in treatment of ailments of immune system and improvement of immunity or as a stimulant (ISD).

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the communities in Montenegrin mountains this plant is used against skin diseases and for the wounds (Menković et al., 2011). The third group of associated variable on the graph pertains to species of Rosaceae, Cornaceae and Crassulaceae applied in form of fresh juice or tisane in cases of immune system disorders and organism strengthening. Namely, numerous responders reported fresh or boiled wild growing fruits such as Pyrus, Malus, Rubus, Rosa and Cornus species, as approved stimulants and health repairing remedy. Finally, appearance of Crassulaceae family on the graph regards the common use of juice extracted from pressed succulent leaves of Sempervivum marmoreum against ear infections and as anti-inflammatory agent. Recent studies suggested that appliance of S. marmoreum might be an important source of natural antioxidants and natural antimicrobial preparations (Stojičević et al., 2008). Again, there were no correlations of any of the tested variables with the gender or age of responders or the geographical position (elevation) of their settlements in the mountain.

3.2. The species most relevant in interviews The frequency of reported medicinal uses for every plant species reported by informants is presented in Table 2. The results of this survey highlighted Hypericum perforatum as the most commonly cited among all reported medicinal species. This species scored the highest number of people interviewed (40.5%) as one of the most commonly indicated herbal drug (including oleum). Among the 12.7% of total indications recorded in the study, indications of H. perforatum preparations mainly include gastrointestinal diseases (peptic ulcer) and immune system strengthening. It is also popular as stimulant, and particularly effective in wound healing according to survey reports. As the chemical composition and pharmacological activities of this plant are well studied, the greatest number of traditional properties was shown to be corresponding with scientific data. The most common use include the aerial part of this plant with inflorescences (Hyperici herba) in the form of infusion, as well as oily extract that is a popular remedy against wounds and burns in folklore medicine on the larger scale. The second most relevant species was Achillea millefolium, reported by 29.7% of informants. The number of indications for this plant scored 8.6% of all uses reported. Most informants apply this plant against genitourinary system disorders (menstrual difficulties, ovarian cysts), nervous system and digestive system disorders or promote other effects of the plant such as appetite stimulant and immune system strengthening. Similarly to the previous plant, its oil is widely used in this region in the form of poultice for healing of purulent wounds. Plantago major shares the same position with previous species taking the equal portion of informants who reported this plant (29.7%), but with a much lower percent of medicinal uses and indication in survey (6.3%). The macerated leaves of this plant are mostly used in the form of poultice in treatment of wounds and

skin ulcers. It is also commonly used in the form of tisane against digestive system disorders and bronchitis. The active principles are generally expected from the leaves, but a few persons apply underground parts or whole plant, which was somewhat unusual. Much less commonly, but also frequently reported species were: Satureja kitaibelii (21.6% informants; 4.1% use reports), Tilia cordata (18.9% informants; 5% use reports), Rubus ulmifolius (16.2% informants; 4.5% use reports) and Urtica dioica (16.2% informants; 4.5% use reports). Particularly interesting in the group of popular species was endemic Satureja kitaibelii which is on country level commonly known as Rtanj tea, a symbol of this mountain. The species is characterized by a small native range, distributed in E Serbia and W Bulgaria, including enclaves in S Carpathians. Our survey has shown that it is used in treatment of respiratory and immune system disorders, against cold and hypotension. To the contrary, its wide popularity on the larger scale was built on uncertified credibility of its effects on sexual potency in men. In any case, enormous popularity of this species attracts many harvesters from remote regions to Rtanj Mt., which remains one of the areas with the richest population in general. Increase of its exploitation is shown by tons of dried aerial parts launched to the market every single year. At the same time native population of this locally distributed plant is faced with threat of overharvesting.

3.3. Ethnobotanical comparison and taxonomic diversity of local medicinal flora In order to compare the ethnobotanical diversity of Rtanj Mt. with the neighboring mountain regions we used data from nine representative ethnobotanical studies. Exclusive of the influence of socio-cultural background as well as different research methods employed, one of the main problems in ethnobotanical comparisons of floristically more or less similar regions was the different sizes of geographical units (Mustafa et al., 2012; Šavikin et al., 2013). The total number of exploited medicinal species recorded in ethnobotanical surveys of several districts of central Balkan Peninsula, with considerable difference in size, is reported in Table 3. As it is shown, distribution of the species number per district varies in a wide range, from 45 enumerated for Rtanj to 207 species traditionally used in the villages of Prokletije massif. These data were used in this study in order to correlate species number with area size, considering the fact that species richness increases as a function of area (Gleason, 1922; Rosenzweig, 1997). In addition to the previously reported comparisons, we have calculated the index of ethnobotanical richness as based on the number of medicinal plant taxa exploited in selected areas that are considerably different in size. The data on species number (as shown in the table) were used in order to estimate relative (α) diversity, considering the effects of geographical factors on insulate areas with highest diversity values. Calculated indices highlighted the Prokletije massif (0.2427) as one of the most important

Table 3 Comparison of medicinal species richness between Rtanj Mt. and different districts of centralBalkan Peninsula according to the previously conducted ethnobotanical studies. District

km2 (A)

Year(s) when the field studies were conducted

No. of medicinal taxa (S)

No. of regionally specific taxa

Species richness (LogS/LogA)

Reference(s)

Prokletije

3500

2004–2010

207

92

0.2427

Kopaonik Gollak Sharr Pešter Rtanj Zlatibor

150 665 840.2 463 296.4 6140

2002–2005 2009 2009, 2011 2010 2011–2012 2011

83 83 76 62 45 69

17 15 21 13 7 10

0.2347 0.2175 0.2107 0.2068 0.1951 0.1879

Pieroni et al. (2005), Pieroni (2008), Menković et al. (2011), Mustafa et al. (2012) Jarić et al. (2007) Mustafa et al. (2012, 2012a) Rehepi et al. (2013) Pieroni et al. (2011) recent study Šavikin et al. (2013)

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district regarding the ethnobotanical richness in central part of the Balkan Peninsula. This fact is in accordance with high floristic diversity of that mountain region, including more than 1600 plant species (Mustafa et al., 2012a), marking it as one of the most important biodiversity centers of the Balkan Peninsula. On the other hand, the results of analyses have indicated that Rtanj Mt. is an area with comparatively low level of ethnobotanical richness (0.1951) among the compared areas. Moreover, the level of ethnobotanical knowledge in this region is decreasing, considering the data on species number reported in the study conducted in the first half of 1970 s by Milojević (1988). This fact could indicate potential loss of ethnobotanical knowledge related to the high emigration rate from the region during the last few decades. From the other side, the lesser level of ethnobotanical knowledge than expected would accurately depict traditional knowledge of a selected group of persons including higher validation of selected species, that was commented by Akerreta et al. (2007) and Šavikin et al. (2013). Many of the recent studies emphasize the level (role) of floristic similarity between the lists of medicinal plants recorded in different ethnic groups or regions of the Balkan Peninsula (Pieroni et al., 2011; Mustafa et al., 2012, 2012a; Šavikin et al., 2013). One of the most exploited methods to determine relation between species list of actual toward other selected areas was by calculating similarity coefficient according to Jaccard (1932). AHC analysis performed using extracted medicinal plants from the species group listed in different ethnobotanical studies has indicated several groups of areas according to the similarity in their floristic composition (Fig. 3). The deep dichotomy in the dendrogram clearly additionally separates Prokletije from the rest of the compared areas that form a distinct and more similar group. Prominent ethno-medicinal heterogeneity of this multiethnic mountain region in comparison to other regions is a reflection of the high number of 92 plant species (or 44.9% of totally reported) which are exclusively exploited in medicinal purposes at that particular locality. It is also based on the largest number (207) of species reported in interviews conducted in that area (Pieroni et al., 2005;Pieroni, 2008; Menković et al., 2011; Mustafa et al., 2012a). Further analyses have indicated presence of two main subclades among the remaining areas of central Balkan Peninsula. One clade united the massifs of Kopaonik and Šara Mt., as both are similar in having comparatively large lists and presence of high-mountain elements of medicinal plants. Including the largest group of compared entities, the last clade of the graph indicates the floristic similarity of four moderately high mountain regions including the study area. According to species composition, the mountain

Fig. 3. Phenogram of ethnobotanical similarity between eleven compared mountain regions of central Balkan Peninsula based on exploited medicinal species.

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of Rtanj is most closely related to Pešter and Zlatibor districts, mostly characterized by dominance of widespread and commonly used medicinal plants. The list of traditionally used species in ethnomedicine of Rtanj region almost completely matched the list from other regions, except for the three endemic species – Achillea clypeolata, Satureja kitaibelii and Thymus jankae – which were not reported in other ethnobotanical surveys. As they are considered local in distribution, they are particularly interesting for future chemical and pharmacological studies. Due to their local distribution, their chemical composition and biological activities are the major reasons for the continuing interest in these species (Kulevanova et al., 1998; Milosavljević et al., 1999; Mihajilov-Krstev et al., 2011). Their importance lies in the possibilities of finding new uses for medicinal plants and discovering new herbal remedies (Menković et al., 2011). In the study region, as well as in whole country, harvesting of medicinal and particularly aromatic plants is a very frequent activity that results in strong negative impact on their natural populations. From the list of over-harvested or illegally collected species in Serbia (Radović and Kozomara, 2011), Centaurium erythraea, Arctostaphylos uva-ursi, Hypericum perforatum, Teucrium montanum and Allium ursinum were recorded in the flora of Rtanj, while Carlina acaulis, Gentiana asclepiadea and Orchis morio populations could be under serious prolonged threat due to harvesting of their underground parts. Some of the exceptionally local representatives are freely used in traditional medicine among the local community from this mountain. Among them a special notion should be given to Nepeta rtanjensis, a strictly endemic species reported exclusively from Rtanj Mountain. Among the inhabitants of the mountain, Nepetae rtanjensis herba is indicated as a remedy for inflammation of the upper respiratory tract (Jančić et al., 1995). Another important aspect of Nepeta rtanjensis is that it is also protected under the CITES convention (CITES, 1973). According to Red List Categories and Criteria (IUCN, 2001), this important plant is also categorized within the list of critically endangered (CR) taxa in the Red Data Book of the flora of Serbia (Stevanović, 1999). The entire population is estimated to about several hundred individuals inhabiting a few micro locations on the mountain, while it is heavily collected by local people due to its strong fragrance (Diklić, 1999).

4. Conclusions This study provided a detailed review of 45 utilized medicinal species representing over 20 families of vascular plants. It includes versatile aspects on their uses in traditional medicine that make Rtanj Mountain interesting from the ethnobotanical point of view and for further scientific and especially phytotherapeutic research. The study of ethnobotanical heritage has shown comparatively strong connections between the local community and natural resources of wild-growing medicinal plants. A specific selection of harvested species, including some endemic and rare representatives, is used among the local people in accordance to the original ethnobotanical practice. High informant consensusfactor value for this region pointed to traditional and specific relatives-chain transport of ethnobotanical information within the analyzed population. Prominent homogeneity and similarity of procedures indicates a collective approach in species choice, uses and treatments by local community. High correlation between harvested plants, methods of preparation and targeted health problems pointed to consistency at least in several cases of their application. Hypericum perforatum was the most relevant species as it was used for the widest spectrum of health problems. The ethnobotanical similarity in the region is further supported by some common phytotherapeutic uses of Veronica officinalis in the

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treatment of skin diseases that happen to be neglected in Serbian traditional medicine. In comparison to the other published studies, comparatively smaller number of medicinal plants is traditionally used. The analyses have shown that, when compared with the other mountain regions of central Balkan Peninsula, the level of ethnobotanical richness of Rtanj Mountain is comparatively low, in contrast to Prokletije region which is the most diverse and with largest potential for medicinal plants. Moreover, the number of conventionally used medicinal plants on Rtanj Mountain has considerably decreased during the last decades, indicating negative trends in ethnobotanical culture. The trends of permanent depopulation have probably caused a partial loss of traditional knowledge. In the future, ethnobotanical heritage should be promoted in a broader sense, getting the special position in conservational as well as tourist and economical opportunities of this region, connected with its biological and floristic diversity.

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