Kevin Cianfaglione

One century later: the folk botanical knowledge of the last remaining Albanians of the upper Reka Valley, Mount Korab, Western Macedonia

BackgroundEthnobotanical surveys of the Western Balkans are important for the cross-cultural study of local plant knowledge and also for obtaining baseline data, which is crucial for fostering future rural development and eco-tourism initiatives in the region. The current ethnobotanical field study was conducted among the last remaining Albanians inhabiting the upper Reka Valley at the base of Mount Korab in the Mavrovo National Park of the Republic of Macedonia.The aims of the study were threefold: 1) to document local knowledge pertaining to plants; 2) to compare these findings with those of an ethnographic account written one century ago and focused on the same territory; and 3) to compare these findings with those of similar field studies previously conducted in other areas of the Balkans.MethodsField research was conducted with all inhabitants of the last four inhabited villages of the upper Reka Valley (n=17). Semi-structured and open interviews were conducted regarding the perception and use of the local flora and cultivated plants.Results and conclusionThe uses of ninety-two plant and fungal taxa were recorded; among the most uncommon uses, the contemporary use of young cooked potato (Solanum tuberosum) leaves and Rumex patientia as a filling for savory pies was documented. Comparison of the data with an ethnographic study conducted one century ago in the same area shows a remarkable resilience of original local plant knowledge, with the only exception of rye, which has today disappeared from the local foodscape. Medicinal plant use reports show important similarities with the ethnobotanical data collected in other Albanian areas, which are largely influenced by South-Slavic cultures.

Resilience at the border: traditional botanical knowledge among Macedonians and Albanians living in Gollobordo, Eastern Albania

BackgroundEthnobotany in South-Eastern Europe is gaining the interest of several scholars and stakeholders, since it is increasingly considered a key point for the re-evaluation of local bio-cultural heritage. The region of Gollobordo, located in Eastern Albania and bordering the Republic of Macedonia, is of particular interest for conducting ethnobiological studies, since it remained relatively isolated for the larger part of the 20th Century and is traditionally inhabited by a majority of ethnic Macedonians and a minority of Albanians (nowadays both sharing the Muslim faith).MethodsAn ethnobotanical survey focused on local food, medicinal, and veterinary plant uses was conducted with 58 participants using open and semi-structured interviews and via participant observation.ResultsWe recorded and identified 115 taxa of vascular plants, which are locally used for food, medicinal, and veterinary purposes (representing 268 total plant reports). The Macedonian Traditional Ecological Knowledge (TEK) was greater than the Albanian TEK, especially in the herbal and ritual domains. This phenomenon may be linked to the long socio-cultural and linguistic isolation of this group during the time when the borders between Albania and the former Yugoslavia were completely closed. Moreover, the unusual current food utilisation of cooked potatoes leaves, still in use nowadays among Macedonians, could represent the side effect of an extreme adaptation that locals underwent over the past century when the introduction of the potato crop made new strategies available for establishing stable settlements around the highest pastures. Additionally, the difference in use of Helichrysum plicatum, which is popular in the local Macedonian folk medicine but absent among Albanians, confirms the particular significance of this taxon as it relates to the yellow colour of its flowers in South Slavic folklore.ConclusionBotanical studies with an ethnographic approach are crucial for understanding patterns of use of plants within given cultures. Importantly, such studies can also allow for analysis of the dynamics of change in these TEK patterns over the time. The results of this study may be important as baseline data set to be used in rural development programs in Gollobordo, aimed at fostering community-based strategies of management of natural resources.

Local knowledge on plants and domestic remedies in the mountain villages of Peshkopia (Eastern Albania)

Ethnobotanical studies in the Balkans are crucial for fostering sustainable rural development in the region and also for investigating the dynamics of change of Traditional Ecological Knowledge (TEK), which has broad-sweeping implications for future biodiversity conservation efforts. A survey of local botanical and medical knowledge and practices was conducted in four mountainous villages of the Peshkopia region in northeast Albania, near the Macedonian border. Snowball sampling techniques were employed to recruit 32 informants for participation in semi-structured interviews regarding the use of the local flora for food, medicinal, veterinary and ritual purposes. The uses of 84 botanical taxa were recorded as well as a number of other folk remedies for the treatment of both humans and livestock. Comparison of the collected data with another ethnobotanical field study recently conducted among Albanians living on the Macedonian side of Mount Korab shows a remarkable divergence in medicinal plant uses, thus confirming the crucial role played by the history of the last century in transforming TEK. Most noteworthy, as a legacy of the Communist period, a relevant number of wild medicinal taxa are still gathered only for trade rather than personal/familial use. This may lead to unsustainable exploitation of certain taxa (i.e. Orchis and Gentiana spp.) and presents some important conservation challenges. Appropriate development and environmental educational frameworks should aim to reconnect local people to the perception of limitation and renewability of botanical resources.

Impact of Extreme Drought and Warming on Survival and Growth Characteristics of Different Provenences of Juvenile Quercus pubescens Willd

In the context of climatic changes, the adaptive potential of organisms toward new environmental challenges is becoming of central interest in modern ecology. Here we investigate the response of growth characteristics and survival in juveniles of four European provenances (Bulgaria, Germany, Hungary and Italy) of Quercus pubescens Willd. to extreme drought and warming in a full factorial common garden experiment. Our results show that extreme durations of drought exceed the tolerance of Quercus pubescens of all provenances. Plants of Italian provenance showed the highest capacity of whole-individual survival, and those of Bulgarian provenance showed the highest maintenance of the apex under extreme drought. Bud bank characteristics showed no general differences among provenances but responded to warming manipulations. The warming effect resulted in a reduction of collar buds and biomass. In Italian individuals it caused an increase of the apical budbank. This is suggestive of a better adaptation for height growth under warming. Our results imply that significant local adaptation of growth characteristics and survival occurs in Quercus pubescens. However, our findings do not support the idea that southern provenances are a general predictor of better plant performance.

Phytochemistry, micromorphology and bioactivities of Ajuga chamaepitys (L.) Schreb. (Lamiaceae, Ajugoideae): Two new harpagide derivatives and an unusual iridoid glycosides pattern.

Ajuga chamaepitys (L.) Schreb, well-known as Camaepitium or Ground Pine, is an annual herb typical of the Mediterranean area accounting several uses in the traditional medicine. In this work we have, analyzed the plant iridoid fraction together with the essential oil composition and study of the plant indumentum. Finally, we assayed the polar extracts and essential oil obtained from the aerial parts for antioxidant activity and cytotoxicity on tumor cells. The analysis of the monoterpene glycosides allowed us to isolate from roots and aerial parts and to structurally elucidate by NMR and MS the following compounds: ajugoside (1), reptoside (2), 8-O-acetylharpagide (3), harpagide (4), 5-O-β-d-glucopyranosyl-harpagide (5), asperulosidic acid (6), deacetyl asperulosidic acid (7) and 5-O-β-d-glucopyranosyl-8-O-acetylharpagide (8), among which 5 and 8 were two new natural products. Chemotaxomic relevance of these constituents was discussed. The chemical analysis of A. chamaepitys essential oil by GC-FID and GC-MS showed ethyl linoleate (13.7%), germacrene D (13.4%), kaurene (8.4%), β-pinene (6.8%), and (E)-phytol (5.3%) as the major volatile components. The micromorphological and histochemical study showed that iridoids and essential oil are mainly produced in the type III capitates and peltate trichomes of leaves and flowers. Biological evaluations of A. chamaepitys polar extracts and essential oil showed that the former were more potent as radical scavengers than the latter. MTT assay revealed that essential oil and ethanolic extracts were moderately cytotoxic on tumor cells with IC50 of 36.88 and 59.24μg/mL on MDA-MB 231 cell line, respectively, and IC50 of 60.48 and 64.12μg/mL on HCT116, respectively.

Polar constituents, protection against reactive oxygen species, and nutritional value of Chinese artichoke (Stachys affinis Bunge)

In the present work, we studied the chemical composition of Chinese artichoke (S. affinis tubers) by analyzing its polar constituents and its macro- and micro- nutrients. A total of nine compounds were isolated from the tuber ethanolic extract and structurally elucidated by Nuclear Magnetic Resonance (NMR) spectroscopy and mass spectrometry (MS). The marker compounds identified were oligosaccharide stachyose and the organic acid, succinic acid, as well as phenylethanoid and iridoid glycosides. The macronutrient profile was dominated by carbohydrates (36.9% dw), whereas potassium (2.36%) was the most abundant micro-nutrient. The tuber ethanolic extract was able to efficiently protect human cells (Caco-2, SHSY-5Y and K562) against t-BHP-induced oxidative damage.

Identification of highly effective antitrypanosomal compounds in essential oils from the Apiaceae family

The Apiaceae family encompasses aromatic plants of economic importance employed in foodstuffs, beverages, perfumery, pharmaceuticals and cosmetics. Apiaceae are rich sources of essential oils because of the wealth of secretory structures (ducts and vittae) they are endowed with. The Apiaceae essential oils are available on an industrial level because of the wide cultivation and disposability of the bulky material from which they are extracted as well as their relatively cheap price. In the fight against protozoal infections, essential oils may represent new therapeutic options. In the present work, we focused on a panel of nine Apiaceae species (Siler montanum, Sison amomum, Echinophora spinosa, Kundmannia sicula, Crithmum maritimum, Helosciadium nodiflorum, Pimpinella anisum, Heracleum sphondylium and Trachyspermum ammi) and their essential oils as a model for the identification of trypanocidal compounds to be used as alternative/integrative therapies in the treatment of Human African trypanosomiasis (HAT) and as starting material for drug design. The evaluation of inhibitory effects of the Apiaceae essential oils against Trypanosoma brucei showed that some of them (E. spinosa, S. amomum, C. maritimum and H. nodiflorum) were active, with EC50 in the range 2.7-10.7 μg/mL. Most of these oils were selective against T. brucei, except the one from C. maritimum that was highly selective against the BALB/3T3 mammalian cells. Testing nine characteristic individual components (α-pinene, sabinene, α-phellandrene, p-cymene, limonene, β-ocimene, γ-terpinene, terpinolene, and myristicin) of these oils, we showed that some of them had much higher selectivity than the oils themselves. Terpinolene was particularly active with an EC50 value of 0.035 μg/mL (0.26 µM) and a selectivity index (SI) of 180. Four other compounds with EC50 in the range 1.0-6.0 μg/mL (7.4-44 µM) had also good SI: α-pinene (>100), β-ocimene (>91), limonene (>18) and sabinene (>17). In conclusion, these results highlight that the essential oils from the Apiaceae family are a reservoir of substances to be used as leading compounds for the development of natural drugs for the treatment of HAT.

Larvicidal Activity of Essential Oils of Five Apiaceae Taxa and Some of Their Main Constituents Against Culex quinquefasciatus

Apiaceae are aromatic herbs producing essential oils which are used on an industrial scale for various purposes. Notably, Apiaceae essential oils may replace synthetic insecticides keeping most of their efficacy and avoiding environmental pollution and human poisoning. In the present work, we explored the insecticidal potential of the essential oils from five Apiaceae taxa, namely Sison amomum, Echinophora spinosa, Heracleum sphondylium subsp. sphondylium, Heracleum sphondylium subsp. ternatum, and Trachyspemum ammi, as well as their major constituents (sabinene, p‐cymene, terpinolene, myristicin, and thymol), against the filariasis vector Culex quinquefasciatus. For the purpose, the essential oils were obtained by hydrodistillation and their composition was achieved by gas chromatography/mass spectrometry (GC/MS). Their acute toxicity on third instar larvae of C. quinquefasciatus was determined. The two most active essential oils were those from T. ammi fruits and E. spinosa roots, showing LC50 below 20 μl/l and LD90 below 50 μl/l. These oils were dominated by the monoterpene phenol thymol and the phenylpropanoid myristicin, respectively, which showed the strongest larvicidal activity (LC50 of 15.1 and 16.3 μl/l, respectively) among the pure compounds tested. These results showed that Apiaceae may be useful as source of larvicidal compounds to be used for the development of cheap, effective and eco‐friendly insecticidal formulations.

Cytotoxic Essential Oils from Eryngium campestre and Eryngium amethystinum (Apiaceae) Growing in Central Italy

Eryngium campestre and E. amethystinum are thorny herbs belonging to the Apiaceae family and spontaneously growing in stony pastures and dry meadows, preferentially on calcareous substrates. In the Mediterranean countries, these plants have been used as a food or traditional remedies to treat various ailments. In the present work, we have analyzed the chemical composition of the essential oils distilled from the aerial parts by GC‐FID and GC/MS, and evaluated their cytotoxic effects on a panel of human cancer cells, namely A375 (human malignant melanoma), MDA‐MB 231 cells (human breast adenocarcinoma), and HCT116 cells (human colon carcinoma), by the MTT assay. Furthermore, the Eryngium essential oils were evaluated for antioxidant and acetylcholinesterase (AChE) activities. The two essential oils were rich in sesquiterpene hydrocarbons, with germacrene D as the major compound, accompanied by allo‐aromadendrene, β‐elemene, spathulenol, and ledol. They turned out to be highly cytotoxic on the tumor cells, with IC50 values (1.65 – 5.32 and 1.57 – 2.99 μg/ml for E. amethystinum and E. campestre, respectively) comparable or close to those of the anticancer drug cisplatin. The E. amethystinum essential oil exhibited a moderate antioxidant activity, whereas that of E. campestre a weak AChE inhibition.

Volatile profiles of flavedo, pulp and seeds in Poncirus trifoliata fruits

BACKGROUND Poncirus trifoliata, also known as trifoliate orange, is a tree native to China and Korea and widely used all over the world as a rootstock breeding material. In this study the differences among the volatile profiles of flavedo, pulp and seeds from two cultivars (var. trifoliata and var. monstrosa) grown in Italy (Marche, Abruzzo and Sicily) were determined. Headspace solid phase microextraction and hydrodistillation techniques were used in combination with GC/FID and GC/MS to obtain the volatile profiles of the samples. RESULTS Both techniques permitted the differentiation of fruit parts based on the main volatile components: the flavedo was characterized by monoterpene hydrocarbons such as limonene and myrcene, the seeds were characterized by sesquiterpene hydrocarbons such as (E)-caryophyllene and germacrene D, while the pulp showed an intermediate composition. The main differences in volatile profiles obtained by the two techniques were analyzed by chemometric techniques such as principal component analysis. CONCLUSION The study did not highlight significant differences in volatiles between the two cultivars of trifoliate orange, while few differences in the number of volatiles in the fruit parts were revealed by the two techniques.