An ethnobotany of the Lukomir Highlanders of Bosnia & Herzegovina

Background This aim of this study is to report upon traditional knowledge and use of wild medicinal plants by the Highlanders of Lukomir, Bjelašnica, Bosnia and Herzegovina (B&H). The Highlanders are an indigenous community of approximately 60 transhumant pastoralist families who speak Bosnian (Bosanski) and inhabit a highly biodiverse region of Europe. This paper adds to the growing record of traditional use of wild plants within isolated communities in the Balkans. Methods An ethnobotanical study using consensus methodology was conducted in Lukomir in Bjelašnica’s mountains and canyons. Field work involved individual semi-structured interviews during which informants described plants, natural product remedies, and preparation methods on field trips, garden tours, while shepherding, or in settings of their choice. Plant use categories were ranked with informant consensus factor and incorporated into a phylogenetic tree. Plants cited were compared to other ethnobotanical surveys of the country. Results Twenty five people were interviewed, resulting in identification of 58 species (including two subspecies) from 35 families, which were cited in 307 medicinal, 40 food, and seven material use reports. Individual plant uses had an average consensus of five and a maximum consensus of 15 out of 25. There were a number of rare and endangered species used as poisons or medicine that are endemic to Flora Europaea and found in Lukomir. Ten species (including subspecies) cited in our research have not previously been reported in the systematic ethnobotanical surveys of medicinal plant use in B&H: (Elymus repens (L.) Gould, Euphorbia myrsinites L., Jovibarba hirta (L.) Opiz, Lilium bosniacum (Beck) Fritsch, Matricaria matricarioides (Less.) Porter ex Britton, Phyllitis scolopendrium (L.) Newman, Rubus saxatilis L., Silene uniflora Roth ssp. glareosa (Jord.) Chater & Walters, Silene uniflora Roth ssp. prostrata (Gaudin) Chater & Walters, Smyrnium perfoliatum L.). New uses not reported in any of the aforementioned systematic surveys were cited for a total of 28 species. Thirteen percent of medicinal plants cited are endemic: Helleborus odorus Waldst. et Kit., Gentiana lutea L., Lilium bosniacum (Beck) Fritsch, Silene uniflora Roth ssp. glareosa (Jord.) Chater & Walters., Silene uniflora Roth ssp. prostrata (Gaudin) Chater & Walters, Salvia officinalis L., Jovibarba hirta (L.) Opiz, and Satureja montana L. Conclusions These results report on the cohesive tradition of medicinal plant use among healers in Lukomir, Bosnia and Herzegovina. This work facilitates the community’s development by facilitating local and international conversations about their traditional medicine and sharing insight for conservation in one of Europe’s most diverse endemic floristic regions, stewarded by one of Europe’s last traditional Highland peoples.


Background
The aim of this study is to report upon traditional knowledge and use of wild medicinal plants by the Highlanders of Lukomir, Bjelala, B, Bosnia and Herzegovina (B&H). The project began as a collaboration with the late ethnobotanical authority in B&H, Professor Sulejman Redžić (1954Redžić ( -2013 and was funded by a post war development grant from the Canadian International Development Agency (CIDA). The first ethnobotanical study published from Lukomir was contributed in Redžić's honour [1].
The project was designed to help describe Lukomir's biocultural region with the goal of assisting B&H to meet environment and health mandates of the European Union Stabilization and Association Agreement.
Lukomir is an isolated village located about 50 km southwest of the capital city, Sarajevo (Fig. 1). Lukomir is situated above Rakitnica canyon with only one access road, which is impassable for part of the year. This isolation has resulted in the preservation of many aspects of traditional lifestyle in Lukomir, including traditional architecture, clothing, herding and medicinal plant gathering and usage [2].
The Lukomir Highlanders are Bosniak and speak Bosnian (Bosanski). They are one of Europe's indigenous communities whose members still occasionally practice a traditional transhumant pastoralist lifestyle. They have inhabited the Lukomir territory (Figs. 1 and 2) within the Dinaric Alps for centuries and are historically Bogomil (Sulejman Redžić, personal communication). Autosomal STR loci studies link Lukomir to the isolated Adriatic islands of Brač, Hvar, and Korčula in Croatia [3]. The Highlanders also have a historic connection to transhumant peoples from the Podveležje Plateau near Mostar, B&H, who travelled to Bjelašnica with their livestock in search of water during the summer months [4].
Until recently, shepherds with hundreds in their flocks set out on grazing journeys throughout the Lukomir territory and to neighbouring regions, especially Konjic, B&H. In the wild, shepherds slept with their sheep flocks while other Lukomir family members gathered and cultivated for enough food and fodder to last the long winter.
Since the Bosnian War (1992-5), many inhabitants have moved to cities and herding has declined. Instead, there is a seasonal migration of former inhabitants to Lukomir to gather medicinal plants, make hay during the summer months, and carry on other traditional practices [2]. According to local informants, the number of people staying in Lukomir all winter and the number of people returning in the summer is declining. In the winter of 2007, three families stayed for the winter. In 2008 and 2009, only two families remained all winter. The winter of 2011-2012 was the first winter without inhabitants. This transition in Lukomir's cultural history was likely exacerbated by 15 m high snowdrifts, an impassable road, and relocations for work and education.

Healthcare in Lukomir
From 1950 onward, the development of the only road prompted the Highlanders to gradually relocate from Donji Lukomir (Lower Lukomir) to their current location of Gornji Lukomir (Upper Lukomir), which is now generally referred to as Lukomir (see DL and GL in Fig. 2). Before this road, Donji Lukomir provided the most efficient access to health and trade services in neighbouring regions via the Rakitnica canyon trails. Lukomir is without a primary healthcare facility, but is known to practice traditional medicine, according to Professor Sulejman Redžić.

Biodiversity in Lukomir
Approximately 60 % of all vascular plant species listed in the Flora Europaea are found in the Balkan Peninsula, making Lukomir part of a highly biodiverse region of Europe [5,6]. Many other studies of traditional plant use for medicine and nutrition have been conducted in the Balkans, including in B&H. Three key systemic ethnobotanical surveys of B&H have recently identified hundreds of plants and thousands of preparations used in traditional human therapy [6][7][8]. Many of these plants are used in official pharmacopeias while others are less known and invite further ethnopharmacological studies. Many of the same uses and preparations were found across regions and ethnic groups, showing an established base of medicinal plant use. This study joins a growing collection of research whose goal is to provide an inventory and increase understanding of medicinal plants used in the Balkan Peninsula [9], including countries such as Albania [10][11][12], Montenegro [13], Kosovo [14], Serbia [15,16], Bulgaria [17,18], and Macedonia [19,20]. An ethnobotanical study using consensus methodology was conducted with the Lukomir Highlanders of B&H in the highlands of Bjelašnica, located southwest of the capital city Sarajevo (Fig. 1). The Lukomir biocultural area is an alpine biogeographic region that is bordered closely by Mediterranean and Continental biogeographic regions within the biocultural area [21]. Many community members are descendants of a Bogomil lineage who first settled in Donji Lukomir (Lower Lukomir) (43.632 lat., 18.194 long., 1200 m a.s.l.) and eventually moved to Gornji Lukomir (Upper Lukomir, commonly referred to as Lukomir) (43.637 lat., 18.182 long., 1460 m a.s.l.) (Fig. 2).

Partnership, permits, and prior informed consent
We visited Lukomir repeatedly, in the summers of 2008 (June, July, August), 2009 (August), 2010 (June), and 2012 (August). By volunteering our help to the Highlanders while shepherding, collecting food, and stacking hay, we had more time for interviews and recruited 25 informants from Lukomir who described plants on mountain and canyon field trips, garden tours, while shepherding, or in settings of their choice. The informants were community healers who ranged in age from younger adults to elders. Fifty-six percent were women. Notes and photos were taken when participants chose to display preparation methods of plant and natural remedies. Plant vouchers, the authors' working field guide, and an iPad (Apple, Cupertino, USA) were used to display collections and photos to elders who could not venture over the mountainside, or for informant review purposes [22].
Land mines were avoided by consulting with a map from the Bosnia and Herzegovina Mine Action Center [23]. Only parts of the Lukomir territory were surveyed by BH MAC and so all plants were collected on trail sides or in areas that were constantly travelled by sheep herds.
While this work was focused on medical plant use, a non-exhaustive record of food and material uses was made in association with the medicinal plants. Due to the expense and time associated with ethnobotanical projects, we felt that including the food and material mentions would be of greater value to the Lukomir community.

Ethnobotanical analysis and statistics Human selection of plants
Consensus on an individual usage was the number of informants using the plant out of 25. For categories of phytochemical and pharmacological interest, the informant consensus factor (F ic ) function, created by Trotter and Logan [24], was used to supply a numerical ranking factor based on the number of use reports (n ur ) and the number of taxa (n taxa ) per usage category.

Statistics and phylogenetic tree
Statistical analysis was conducted using Prism 6 software. A phylogenetic tree was constructed using TreeGraph 2.0.47-206 beta, FigTree v1.4.0 (Fig. 3). Taxa are based on the flora of B&H [25] and corrected to match accepted names in Tropicos [26] and the topology presented is based on Angiosperm Phylogeny Group III [27].

Results and discussion
Interviews During our interviews, the Highlanders identified 58 species (including 2 subspecies) from 35 families, which were cited in 307 medicinal, 40 food, and seven material use reports and are included Table 1. Grassland habitats generated the most use reports of plants. Medicinal plant collection was greatest from mid-July to early August as many healers collected plants while in the grasslands harvesting hay. The average consensus on use of medicinal plants was 3.5 informants out of 25, and a maximum consensus of 15 was obtained on the medicinal plants Mentha longifolia (L.) L. and Salvia officinalis L. Average food plant consensus was three. The maximum food plant consensus was eight for Urtica dioica L. Finally, there was a maximum consensus of two for material plants Pinus nigra Arnold and Cornus mas L.
For a community of approximately 60 households, almost half of the households were interviewed. We wondered if the list of medicinal plants was small; however, when looking at our cross-cultural comparison of Cree, Highlanders and Maya we noted that while Lukomir had fewer use reports than the Maya, they had much more than the Cree [28]. This made sense to us that the Cree had less than Lukomir because of their northern location, less biodiversity and cultural assimilation via residential schools in Canada, and that Lukomir healers had fewer reports than the Maya because the Maya live in a tropical rain forest with much greater biodiversity. Lukomir's data, within a continental, alpine and Mediterranean geography in the middle of these two extremes felt reasonable to us.

Frequency of use reports per usage category
The number of use reports per usage category (Fig. 4) varied from 1 to 72. High frequency categories included: genitourinary system disorders, panacea, pain,    and circulatory system disorders. Medium frequency categories included: food, skin/subcutaneous cellular tissue disorders, respiratory system disorders, and illdefined symptoms with many low frequency results (Fig. 4). The high use of plants for genitourinary system disorders reported (20 %), especially gynaecological uses, may be linked to researcher LŠ and the majority of interviewees being female (n = 14 of 25, 56 %), as women would be more likely to have knowledge or experience of these uses and may also have been more comfortable sharing this information with a woman interviewer. However, high female input was predicted based on our survey of the literature [29].
Panacea treatments were highly cited (14 %) since they were often cross-listed as food plants, which included a number of economically important plants. High requests for panacea treatments is probably linked to diabetes, which the visiting physician cited as a top health concern. Diabetic insults affect many tissues and organs, leading to symptoms like frequent urination, neuropathy, and slow healing sores, and panacea treatments are an important resource for Highlanders with these symptoms.
Pain treatment was a high frequency usage (13 %), likely due to hard labor in wet and cold conditions. This may also be linked to heart disease which was another top health concern described by the visiting physician. These high frequency categories indicate that Highlander community members are requesting traditional medicines in line with major health concerns.

Frequency of taxa per usage category
Frequency of taxa per usage category distributions revealed the type of usages of most concern (Fig. 5). The medicinal categories employing the largest number of taxa included genitourinary system disorders, pain, and skin/subcutaneous cellular tissue disorders, indicating that these categories are of high community concern. Medium categories included food, panacea, circulatory system disorders, ill-defined symptoms, respiratory system disorders, and infections/infestations and may mark chronic or recurring health conditions that present more often than low frequency categories (four species or less). Table 2 separates the number of taxa into specific uses in their associated usage category.
High diversity (five or more taxa) was found in the following uses: panacea, internal medicine, heart trouble, genitourinary system health, stomach pain, cuts, and infections.

Informant consensus factor (F ic ) per usage category
For a disease level analysis, the F ic values were calculated (Fig. 6)   culture) and 0.08 (Idate culture) [30]. The very low value for Idate culture may be due to their culture of secrecy surrounding medicinal plant use. Both Lukomir averages were also greater than averages calculated from a study by Amiguet et al. [31], with the Q'eqchi' Maya of Belize (average F ic = 0.62), and the study with the Yucatec Maya of Mexico (F ic = 0.48) [32]. This difference in F ic results may be explained by recognizing that the Maya informants were from isolated villages while the Lukomir Highlanders, although transhumant, share a single village where they traditionally stayed all winter, snow-locked, and where the interviews were conducted.
These results indicate that these plants are well known in a well defined community-based traditional knowledge system. When consensus is high, these categories are more likely to be active in condition-related pharmacological assays [33].

Analysis of medicinal plant families
As a first step to identifying key plant families, the number of medicinal, food, and material species by family was ranked (Fig. 7). The three most diverse families were the Lamiaceae (7 species), Asteraceae (6), and Rosaceae (5).
Four medicinal taxa were cross listed as foods, showing that medicinal and food plant categories are not considered mutually exclusive. These traditional foods are often cited for having disease preventing properties and the loss of traditional diets is associated with the rise of chronic conditions such as heart disease and diabetes [1,34].
The phylogeny (Fig. 3) links plant families to their use reports. Among the top three families, Lamiaceae had 80 use reports (77 medicinal) ( Table 1). The Lamiaceae in Lukomir flourish in the Mediterranean environment on the south-facing dolomite slopes of the Raktinica Canyon. The Lukomir cultural area is a refuge of wild endemic Lamiaceae taxa, as indicated by our ongoing floristic survey. The Caryophyllaceae family also had high use (36 use reports, all medicinal) ( Table 1) and is often found fringing village trails and grasslands. Asteraceae received 32 (30 medicinal) use reports (Table 1), which is not surprising given the family's cosmopolitan distribution throughout Lukomir's alpine, Mediterranean, and continental biogeographical regions.

Traditional medicinal plant preparation
Leaves are the plant part cited most often by the Highlanders (Fig. 8). Infusions were the most common preparation (Fig. 9), as was found in several other studies of B&H [6][7][8]). Some taxa were included in family čaj (infusion) collections, not necessarily viewed as medicine but consumed after mealtime for their general health benefits [1].

Plant usage compared to other studies in Bosnia & Herzegovina
In comparison with other ethnobotanical surveys of medicinal plants used in B&H [6][7][8]) ten species (including subspecies) in our research have not previously been reported (other than in our previous publication from the same project [1] (Table 3). New medicinal uses not reported in any of the aforementioned systematic surveys were cited for a total of 29 species, including the ten newly reported species.

Use of rare and endemic plants
This study considers endemics in the context of Flora Europaea and did not consider garden plants as endemics. Garden plants listed in our survey, such as potato, have been denoted in Table 1. It was observed that Lukomir is a wild refuge for many european endemics. For example, sage was never found in the garden, and was only located once, halfway down Europe's largest canyon.
The Highlanders uses the root of Gentiana lutea L. have not been reported in other studies in the country (Table 1).
Lilium bosniacum (Beck) Fritsch, the Bosnian Lily, is endemic to the central Dinaric Alps whose taxonomic status has been much debated [35]. Studies have come to differing conclusions as to whether L. bosniacum is a distinct species, a subspecies or a variety of L. carniolicum [35][36][37]). Recent molecular cytogenic studies have found evidence to support L. bosniacum's status as a distinct species [35,37]. According to Tropicos, Lilium bosniacum (Beck) Fritsch has species rank [38].   Table 1 Fig . 9 Plant preparation reports of the plant species listed in Table 1  Lilium bosniacum has been classified as a rare and vulnerable taxon in the pending edition of the Red Book of B&H. Despite not being its typical habitat, L. bosniacum is found in Bjelašnica [35]. One informant in Lukomir cited two uses for L. bosniacum.
Salvia officinalis L. (Kadulja, sage) was not found in gardens, but growing wild. In nearby Albania, this plant is an important export product to Western Europe and the United States [39]. This plant was cited a large number of times in the 2007 systemic study of the country's plant use [8].