Wild food plants in Graecanic communities in

Calabria, Southern Italy - Ethnobotany,

current role in Mediterranean diets,

and antioxidant activity

Thesis presented by

Sabine M. Nebel

for the degree of Doctor of Philosophy

Centre for Pharmacognosy and Phytotherapy

The School of Pharmacy

University of London

2006

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London between 2002 and 2006 under the supervision of Prof. Michael Heinrich. I certify that the research described is original and that any parts of the work that have been conducted by collaboration are clearly indicated. I also certify that I have written all the text herein and have clearly indicated by suitable citation any part of this dissertation that has already appeared in publication.

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Signature Date Abstract

Dietary patterns are changing rapidly all over the world. The body of available local food knowledge, which forms the basis of many local traditions, is decreasing dramatically. In rural areas throughout the Mediterranean, vegetables and salads made from wild plants have been particularly important as local foods since ancient times.

However, very little is known about the use of these wild food plants (WFPs) and about their contribution to health, while numerous clinical and pharmacological-biochemical studies have shown beneficial effects for major components of Mediterranean diets.

This research project studied WFPs used in the Graecanic area in Calabria, Southern

Italy as components of the local diet with potential antioxidant activity. It was embedded in an EU-fimded project entitled “Local Food - Nutraceuticals”. The research approach adopted for this interdisciplinary research combined ethnobotanical methods with pharmacology and nutritional sciences.

The gathering, processing and consumption of these plants were studied using participant observation techniques and semi-structured interviews. Local perceptions about WFPs (beneficial health effects, health risks) and nutritional data were obtained through a socio-nutritional study conducted in Italy, Spain and Greece. More than 40

WFPs are used as condiments, or vegetables, including edible greens, called ta chorta in the local language. Many are considered to be healthy because of their bitterness (e.g.

Reseda alba). Furthermore, the antioxidant activity of WFPs from Italy, Spain and

Greece, was evaluated using in vitro assays. Crude extracts were tested for free radical scavenging activity (FRSA), and for the inhibition of xanthine oxidase (XO). Only a small number of plant extracts showed significant antioxidant activity. However, some extracts had promising activity in the XO-system.

This interdisciplinary research contributed not only to the understanding of locally used

WFPs as a promising source of natural antioxidants, but also to the safeguarding of this rapidly vanishing local knowledge. éla condàmu ce dommu to chéri vienimi vicino e dammi la mano a ttelise i cardîamu na mi pesàni se vuoi che il mio cuore non muoia i agapimîa e(ne) fola ena lulùdi l’amore è come un fiore an den ndo potise spisio ecino cottèni se non lo innaffi spesso esso appassisce

1 zoi ene mien la vita è breve éla agàpimu vieni amor mio ti ego de ssonno zi che io non posso vivere

Mimmo Nucera Milinari di Gallicianô 6

Acknowledgements

I wish to express my sincerest thanks to:

- the inhabitants of the Graecanic area in Calabria, particularly the inhabitants of

Gallicianô, for their enthusiasm for this project and willingness to share their knowledge with me, and their lovely hospitality: they have given me a second home. Furthermore, I would like to thank all the members of the Cumelca Association for their support throughout my stay in Gallicianô - 1 still feel like dancing whenever I hear la tarantella', the Mayor of Condofuri, Nucera Giovanni S. and the town councillor, Manti Natale and for the logistic support.

- to Prof. Michael Heinrich as my supervisor, to Dr. Jo Barnes (second supervisor). Dr.

Andrea Pieroni, Dr. Marco Leonti, Prof. Felicity Smith and Dr. José Prieto-Garcia from the School of Pharmacy for their scientific support. Thanks also to Dr. Cristina

Inocencio Pretel and Rafael Llorach from the University of Murcia, Spain; to Dr.

Zaxarias Kypriotakis (Exoli gsoponias, TEI Kritis, Crete, Greece), Dr. Manuel Pardo de

Santayana (Royal Botanic Garden in Madrid), Andreas Lardos (Switzerland) for discussions about specific aspects of the project, as well as to all members of the consortium “Local Food-Nutraceuticals”. Furthermore, I would like to thank Dr.

Nicholas Hind from the Royal Botanical Gardens, Kew, UK, for his help with the identification of some Asteraceae species. Thanks also to the Erasmus students Sandra

Maier, Lydia Tomantschger and Henrik Nilsson for their contribution to this thesis.

- to all my colleagues and staff at the Centre for Pharmacognosy and Phytotherapy,

School of Pharmacy, University of London, UK.

- last but not least I express my deepest thank to Caspar Schmidt, and to both our families for their love, support and patience. Financial support during the work of this thesis was obtained from:

The School of Pharmacy, University of London,

The British Federation of Woman Graduates (BFWG)

- The European Commission, DG Research [Quality of Life and Management of

Living Resources, key action 1 : Food, Nutrition and Flealth (“Local Food-

Nutraceuticals”, contract number: QLKl-CT-2001-00173, coordinator: M. Heinrich Table of Contents

Abstract...... 3 List of Tables...... 11 List of Figures...... 13 Abbreviations...... 15

1 Introduction...... 17

1.1 Local food as an element of traditional knowledge - a theoretical perspective...... 21

1.2 Local foods of the Mediterranean...... 26

1.3 The changing food pattern...... 30

1.4 Food as medicine - medicine as food...... 32

1.5 The *Tocal Food-Nutraceuticals” project and its goals...... 34

2 Wild food plants used in the Graecanic area in Calabria, Southern Italy 37

2.1 Introduction...... 38

2.1.1 The ethnographic background: Graecanic communities in Calabria, 81... 40 2.1.2 Historical sources of botanical data ...... 43

2.2 Aims and Objectives...... 45

2.3 Methods...... 46

2.3.1 Ethnobotanical methods ...... 47 2.3.2 Historical-linguistic method ...... 49

2.4 Results and Discussion...... 50

2.4.1 Wild food plants traditionally consumed in the Graecanic area ...... 50 2.4.2 Traditional dishes made of wild food plants ...... 66 2.4.3 Historical importance of wild food plants ...... 69 2.4.4 Food as medicines ...... 75 2.4.5 Dissemination of ethnobotanical information ...... 77

2.5 Conclusion...... 80 3 Socio-nutritional study in six rural areas of the Mediterranean...... 83

3.1 Introduction...... 84

3.2 Aims and Objectives...... 86

3.3 Methodology ...... 87

3.3.1 Sampling and recruitment ...... 88 3.3.2 Data collection ...... 91 3.3.3 Data processing and analysis ...... 96

3.4 Results and Discussion...... 97

3.4.1 Recruitment settings: households, response rate, and interviewers ...... 97 3.4.2 Characteristics of study sites and respondents ...... 98 3.4.3 Wild food plant consumption ...... 102 3.4.4 Perception regarding health benefits or risks ...... 108 3.4.5 Factors influencing the choice of food products ...... 110 3.4.6 General aspects of lifestyle and health ...... I l l 3.4.7 Questionnaire surveys: Methodological problems for field-based ethno- pharmacologists ...... 116

3.5 Conclusion...... 119

4 Antioxidant activity of wild food plants ...... 122

4.1 Introduction...... 123

4.1.1 Health beneficial effects of the Mediterranean diet ...... 123 4.1.2 Oxidative stress and natural antioxidants ...... 124

4.2 Aims and Objectives...... 129

4.3 Materials and methods...... 129

4.3.1 Collection of plant material ...... 129 4.3.2 Preparation of crude plant extract ...... 130 4.3.3 Antioxidant assay: measurement of free radical scavenging activity (FRSA) in the 1,1 -diphenyl-2-picrylhydrazil radical (DPPH) assay ...... 131 4.3.4 Enzyme inhibition assay: Inhibition of xanthine oxidase ...... 132 4.3.5 Further assays part of the LFN analysis ...... 133 10

4.3.6 Phytochemistry of Reseda alba L...... 134

4.4 Results and Discussion...... 134

4.4.1 Antioxidant activity ...... 140 4.4.2 Enzyme inhibition activity ...... 141 4.4.3 Results of fiirther assays part of the LFN analysis ...... 142 4.4.4 Phytochemistry of Reseda alba L...... 146

4.5 Conclusion...... 147

5 Conclusion...... 149

6 References ...... 152

7 Publications ...... 185

8 Appendix ...... 188

8.1 Ethnobotanical questionnaire (in Italian)...... 189

8.2 Socio-nutritional questionnaire (in Italian)...... 193

8.3 List of wild food plants used in the primary screening (Antioxidant activity) ...... 208

8.4 CD *^ta chorta: Piante commestibili tradizionali di Gallicianô’’...... 213 11

List of Tables

Table 1: Non-cultivated (wild) food plants traditionally gathered and consumed in the

Graecanic communities in Calabria (Gallicianô and other villages) ...... 51

Table 2: The most frequently mentioned wild food species in Gallicianô, Graecanic area,

Calabria, Southern Italy ...... 63

Table 3: Examples of local dishes with WFPs in the Graecanic area in Calabria,

Southern Italy ...... 68

Table 4: Cognates of chorta (edible greens) and other WFPs used in Gallicianô,

Calabria, Southern Italy and in Greece ...... 70

Table 5: Food plants used as medicine in Gallicianô, Calabria, Southern Italy ...... 75

Table 6: Overview table of number of inhabitants and households in the selected study

sites of the Mediterranean ...... 89

Table 7: The structure of the socio-nutritional questionnaire used in the “Local food —

Nutraceuticals” project ...... 91

Table 8: Sample size (number of households included in the study), total of interviews

conducted, response rate in all six study areas and the interviewers ...... 98

Table 9: Current employment status of household members in the six study areas 101

Table 10: Sex, Age of respondents in the six study areas and analysis of responsibilities

for deciding what foods are eaten in the household ...... 102

Table 11 : Consumption of wild food plants (results of the FFQ) linked with the

frequency of wild food plant consumption (section 1 of the questionnaire). 103

Table 12: Total consumption of fruits and vegetables (including WFPs) per week and

per day ...... 107 12

Table 13: Percentage of individuals reporting problems on the EQ-5D descriptive

system: the mean VAS by study sites...... 113

Table 14: Diseases diagnosed/ treated in the past year in the three countries (gender

specific) ...... 115

Table 15: Assays conducted by the LFN partners (The Local Food-Nutraceuticals

Consortium 2005) ...... 133

Table 16: Definition of activity range (% of control) for the performed bioassays ...... 136

Table 17: Summary of primary screening data ...... 137 13

List of Figures

Figure 1: Model of Biodiversity, Diet and Health in the context of global change

(adapted from Johns 2001) ...... 34

Figure 2: Map of Italy ...... 41

Figure 3: Graecanic area in the Province of Reggio di Calabria, Region of Calabria.

Graecanic villages: Amendolea, Bova, Condofuri, Gallicianô, Roccaforte del

Greco and Roghudi ...... 41

Figure 4: Shepherd with flock of sheep and goats ...... 43

Figure 5: Drawing of Marathron - Fennel - Foeniculum vulgare ...... 44

Figure 6: Gallicianô (municipality of Condofuri, RC), in the southern part of the

Aspromonte mountain, 621 metres above sea level, approximately 120

inhabitants ...... 47

Figure 7: Signora Tota and the author, Sabine Nebel during an interview ...... 48

Figure 8: Signora Angela, gathering sculimbri - Leaf stalks of basal leaves of Scolymus

hispanicus are prepared to be cooked with home made pasta {Sculimbri me ta

lasagne) or together with other wild greens ...... 57

Figure 9: The sourrouding area of Gallicianô: Olive groves (uliveto), homegardens

(orto), vineyards {ambeli)...... 59

Figure 10: Bucket with wild gathered ta chdrta (wild greens) ...... 60

Figure 11: Young leaves of Pricaddhida (Lactuca viminea, Asteraceae)...... 62

Figure 12: Inflorescence of gattinaria {Reseda alba, Resedaceae)...... 62

Figure 13: gaddhazzfda {Reichardiapicroides, Asteraceae)...... 62

Figure 14: Chôrta vramena - mixed wild greens {chorta mimmena) boiled ...... 67 14

Figure 15: Picture of “Fasùli me ta mâsara == Beans with wild fennel”. Soup with broad

beans, young leaves of wild fennel, carrots, potato, pasta and fresh chili 67

Figure 16: Booklet “ta chorta: Piante commestibili tradizionali di Gallicianô” by Nebel,

Sabine (2005) School of Pharmacy, University of London, London UK 78

Figure 17: Example of plant monograph from the booklet ‘7a chorta""...... 79

Figure 18: Example of two recipes: “Chorta vramena” and Chorta tiganimena” 79

Figure 19: Box plot of the age distribution in the six study areas (Southern Italy, South-

Eastern Spain and Crete) ...... 99

Figure 20: Perception of WFPs regarding any helpful/beneficial effect on health by local

people in the six study areas in the Mediterranean ...... 109

Figure 21: Signora Maria collecting olives (white bucket) and wild greens (blue bucket)

in her olive grove, Gallicianô, Southern Italy ...... 112

Figure 22: Girls carrying oranges from the homegardens back to the village ...... 112

Figure 23: Percentage of responders rating their health on a scale (excellent to poor) .114

Figure 24: DPPH-H is a product of the reaction between DPPH* and an antioxidant ..127

Figure 25: Xanthine Oxidase (XO) generating superoxid anions (0% ") and hydrogen

peroxide (H 2 O2 ) with the formation of uric acid as the final product 128

Figure 26: Drying edible parts of WFPs collected in spring 2002 on an old bed frame, at

room temperature in Gallicianô, Southern Italy ...... 130

Figure 27: Graph showing the low correlation (R^ = 0.16) between the Total Activity

Score (TAS) and the polyphenol content (PP) of all plant samples 145

Figure 28: kaempferol-3,7-0-a-L-dirhamnoside (kaempferitrin) ...... 146 15

Abbreviations

AChE Inhibition of acetylcholine esterase assay

ARI’s Ageing related illnesses

BrdU Determination of endothelial cell proliferation

Comet Measurement of plant extracts’ protective ability against hydrogen

peroxide-induced DNA damage ct cytotoxic

CT Chorta tiganimena

CV Chorta vramena

CVD Cardiovascular disease

DMSO Dimethyl Sulfoxide

DPPH 1,1 -diphenyl-2-picrylhydrazil

E Q -5 D EuroQol: established tool to measure the health state of individuals

FRSA free radical scavenging activity

HRQoL Health related quality of life hSERT Serotonin re-uptake inhibition

LFN Local Food - Nutraceutical

MDA Measurement of lipid peroxidation in mouse brain tissue

MS Mass spectrometry

Nitrite Inhibition of cytokine-induced cell activation

NMR Nuclear Magnetic Resonance

OxyHb Measurement of hypochlorous acid-induced oxyhaemoglobin bleaching

PP Polyphenol content 16

Abbreviations (continued)

PPARy PPARy binding assay

PTLC Preparative thin layer chromatography

TAS Total activity score

TEK Traditional ecological knowledge

TFK Traditional food knowledge

TK Traditional knowledge

TLC Thin layer chromatography

VAS Visual Analogue Scale

Viability Cytotoxicity assessment

VLC Vacuum Liquid Chromatography

WFPs Wild food plants

XO Xanthine oxidase Chapter 1 Introduction 17

1 Introduction

The Mediterranean basin has a long and multifaceted cultural history and high biodiversity (Hammer et al. 1987; Medail & Quezel 1997). This biodiversity has provided the basis for the development of many agricultural crops, and therefore the

Mediterranean area belongs to one of the primary centres of cultivated plant origin and diversity (Harlan 1971; Heywood 1999a). Also many wild relatives of crops are present in the area (Pignatti 1971) and since antiquity humans have used this diversity of non­ cultivated plants, also commonly referred to as “wild” plants in botanical literature

(Tardio et al 2006). These wild plants are used for many different purposes, such as food, which is the focus of the study in hand, or as medicines for humans and animals, fuel, domestic appliances, in handicrafts or rituals, or for many other purposes (Harlan

1975; Galt & Galt 1978; Heywood 1999b).

Generally, the documentation of food plants in the Mediterranean has a long tradition and goes back to Ancient Greek and Roman times (Trichopoulou & Lagiou 1997). The philosopher Plato, in his “Republic”, gives an example of a healthy diet, including olive oil, cereals, fruits and “country dishes of wild roots and vegetables” (Ferrari 2000, p.

55). Also Apicius included some recipes with local and exotic food plants in the only surviving Roman cookbook (Apicius 1991). However, modem ethnobotanical and ethnopharmaceutical surveys in Southern Europe have mostly dealt with popular phytotherapeutical remedies (e.g. Leporatti & Pavesi 1989; Rivera & Obon 1995; Ertug

2000; El-Hilaly et al 2003) and up to a few years ago very little attention had been paid to wild gathered fruits and vegetables (Forbes 1976; Paoletti et a l 1995; Heywood & Chapter 1 Introduction 18

Skoula 1997; Bonet & Valles 2002; Pieroni et al. 2002b; Guarrera 2003; Ertug 2004;

Pardo de Santayana et al. 2005b; Pieroni et al. 2005; Rivera et al. 2005; Tardio et al.

2006) despite the diversity of traditional food knowledge around the Mediterranean.

Some regions still lack ethnobotanical investigations. Therefore systematic investigations of traditional food knowledge are urgently required in Southern Europe, especially in those areas which, for geographical and historical reasons remain relatively isolated, and where industrial development has not led to a complete decline of the local traditions. The term “traditional” is used in this thesis for defining something that has been an integrated part of a culture for more than one generation (Ogoye-Ndegwa &

Aagaard-Hansen 2003).

The interest in local plant knowledge and practice encompasses the study and analysis of plant species within the local socio-cultural context of the communities involved. Such an approach encompasses a dynamic perspective of studying different patterns of knowledge and practice concerning wild plants as a process of interaction between

“local” and “global” knowledge systems. Ethnobotanical studies among non-dominant ethnic groups in Europe have only been rarely carried out; examples are the surveys on

Catalan ethnobotany in Spain (Raja et al. 1997; Bonet & Valles 2002), a preliminary work on the folk pharmacobotany among Ladin groups in North-western Italy (Barbini et al. 1999) and another ethnobotanical survey among Arbëreshë groups in Southern

Italy (Pieroni et al. 2002b). A research focus on ethnobotany of ethnic, or linguistic minorities in the Mediterranean, as well as small isolated conununities permits us to “re­ discover” retained traditional knowledge, and to try to connect this evaluation with sustaining the biological and cultural/linguistic diversity. Chapter 1 Introduction 1 9

However, traditional knowledge regarding food use is under pressure as dietary patterns change rapidly all over the world (Haddad 2003; Popkin 2004) and the body of available local food knowledge, which forms the basis of many local traditions, is decreasing dramatically. However, assessing changes in local diets is difficult since in many instances “traditional” or local dietary patterns have already disappeared and often no quantitative data on food consumption in traditional societies are available (Tumino,

Frasca, Giurdanella, Lauria, & Krogh, 2002). The ethnobotanical studies related to wild gathered food plants which have been conducted in the Mediterranean so far clearly lack a quantitative approach which would indicate the role and importance of the different wild food plants (WFPs) in traditional diets. Moreover, in recent decades it has become accepted that food and medicine are closely related (Etkin 1996). Plants may be used both as food and as medicine and it is sometimes difficult to draw a line between the two groups. For example, many studies on potential health benefiting aspects of traditional foods show that such plants have specific pharmacological effects (Keys & Keys 1975;

Trichopoulou et al. 1999b). But so far only a few ethnobotanical investigations have included a scientific pharmacological study (Pieroni et a l 2002a) or phytochemical investigations on WFPs. Consequently, both socio-nutritional studies regarding traditional food culture and pharmacological and phytochemical investigations of WFPs are urgently needed.

The starting point of this project was the initial hypothesis; '‘''Wild gathered food plants are still an important element of local dietary patterns in rural areas of the

Mediterranean and they contribute to a healthy and balanced local diet. New leads in the search for nutraceuticals might be found among such wild gathered food plants. ” Chapter 1 Introduction 20

Therefore the objective of this study was to investigate the traditional food of one ethnic and linguistic minority in Southern Italy from an ethnobotanical, socio-nutritional and pharmacological perspective. The study was conducted in Graecanic communities, located in the southern part of the province of Reggio Calabria in the region of Calabria.

The reasons why the Graecanic area has been chosen for this investigation are threefold;

(i) so far no ethnobotanical research has been carried out in this region; (ii) the

Graecanic area was very isolated until the 1950s, when access roads to the villages were built, and therefore they retained a traditional lifestyle until today; (iii) it allows a very interesting intercultural and historical comparison of the ethnobotanical data on local foods with mainland Greece, because the Graecanic communities are of Ancient Greek origin, and have not been in direct contact for many centuries and they live in similar ecological environments.

To test the initial hypothesis, the interdisciplinary research approach adopted for this project combined ethnobotanical methods with pharmacology and nutritional science.

This included the identification of wild food plant species in the selected communities and the documentation of the gathering, processing, cooking as well as the consumption of the different local food plant species. Furthermore, the cultural and historical importance of local food plants was investigated using ethnobotanical and historical- linguistic methods. The role of WFPs in the local diet was assessed by means of a socio- nutritional study. And finally, some health promoting properties of the collected wild food plants were evaluated with different antioxidant assays. According to the results of the primary screening and further criteria, one wild food plant from the Graecanic cuisine was selected for some general phytochemical investigation. This research Chapter 1 Introduction 2 1 contributes not only to the understanding of locally used food plants and potentially to the development of novel health food supplements and novel crops, but also to the safeguarding of these rapidly disappearing local and regional systems of plant-related knowledge and practice of local communities in the Mediterranean Region (cultural diversity).

1.1 Local food as an element of traditional knowledge - a theoretical perspective

In order to better understand the concept of local food it is indispensable to look at knowledge systems from a theoretical perspective.

Knowledge and bearers of knowledge stand at the centre of knowledge systems, which can be defined as “the way people understand the world, and interpret and apply meaning to their experiences” (Wiersum 2000). Such knowledge is built through the complex process of selecting, rejecting, creating, and transforming information, and is inextricably linked to the social, environmental, economical and institutional contexts in which it occurs (Acre & Long 1992). Numerous terms have been used for it and in the following, the focus is on two which are particularly relevant in the context of this thesis: local knowledge and traditional ecological knowledge (TEK). There are other terms, such as traditional knowledge or indigenous knowledge^ which are closely related, partly overlapping, or even synonymous with local knowledge (FAO 2004).

The example chosen is the knowledge about food and how this knowledge impacts on the utilisation of botanical resources in some regions of the Mediterranean. Even though the different social science disciplines have all contributed to an understanding of the Chapter 1 Introduction 22 social aspects of food consumption, hardly any discipline matches anthropology in contributing to our understanding of food consumption, perhaps the classical and most frequently cited example being the culinary triangle of Levi- Strauss (Lévi-Strauss

1968).

Local traditions rely on information being passed on from one generation to the next in one community or in a small region. Food may form a part of local knowledge if its use is confined to a certain area for which it is a characteristic cultural trait. Traditional food knowledge (TFK) as part of local knowledge is strongly influenced and determined by socio-economic and cultural determinants, religion and history. The concept of local knowledge has been essential in the field of anthropology. Originally developed in the context of cultural anthropology in the USA, it is now recognised as an important theoretical vision of how to interpret knowledge systems, which often have not been recognised formally. The contributions by Clifford Geertz (esp. Geertz 1983) have been crucial. His collection of essays “Local Knowledge - Further Essays in Interpretative

Anthropology” clearly is a milestone in the discussion about how people interact with each other and their environment. C. Geertz is one of the co-founders of symbolic or interpretative anthropology, which developed in the late 1960s. Culture is - according to

Geertz - a complex system of symbols, which allows humans to give meaning to the experiences they face and to communicate these experiences among each other. This culture-specific system of symbols should be recorded as a “thick description”, a term he had introduced into the field in 1973 (borrowing it from the British philosopher G.

Ryle). It is essential to understand these systems of symbols and to properly describe them in detail. Thus a “thick description” is one based on the experience of a researcher Chapter 1 Introduction 23 who comes from a foreign culture, which simultaneously shows meaning systems which have not been visible to the indigenous groups. Geertz hoped that the deeper understanding of signs opens and/or increases the dialogue among different cultures

(Geertz 1973).

The symbolic role of “local knowledge” is essential to Geertz's thought and has had a strong impact on anthropological thought. In the first instance it is important to point out that “local knowledge” is an all-encompassing concept. As Geertz (1983, p. 167) has put it focusing on legal questions: “Like sailing, gardening, politics, and poetry, law and ethnography are crafts of place: they work by the light of local knowledge”. Thus all aspects of culture rely on a complex system of thought which in its totality forms local knowledge. Therefore, local food knowledge or any other aspect of local knowledge is just one part of this complex system, which has an important meaning to the members of the culture.

These concepts do provide a strong theoretical foundation for working on the complex meaning systems of indigenous and “traditional” societies. However, they clearly lack a systematic and detailed discussion of applied aspects o f such knowledge systems. There is no input from applied researchers or from anyone who is interested in preserving, disseminating or using this information. Therefore, it is essential to highlight another stream of thought, which does come specifically from such an angle - the concept of

Traditional Ecological Knowledge, which is also well known under its acronym TEK.

This has been defined as “a body of knowledge built up by a group of people through generations of living in close contact with nature” (Johnson 1992). Another definition Chapter 1 Introduction 24 describes TEK as a “cumulative body of knowledge practice and belief evolving by adaptive processes and handed down through generations by cultural transmission, about the relationship of living beings (including humans) with one another and with their environment” (Berkes et al. 2000). Both definitions recognise that TEK is an attribute of societies in general with historical continuity in resource use practice. Interest in TEK has been growing in recent years, partly due to the recognition that such knowledge can contribute to the conservation of biodiversity (Gadgil et at. 1993; Slikkerveer 1999), and to sustainable use in general (Berkes 1999). The understanding of the close relationship between biodiversity and cultural diversity strengthen the sustainable management and use and conservation of plants (concept of bio-cultural diversity) (Maffi 2001).

TEK includes a system of classification, a set of empirical observations about the local environment, a system of self-management that covers resource use. The quantity and quality of TEK varies among community members, depending upon gender, age, social status, intellectual capability and profession (hunter, spiritual leader, healer, etc). With its roots firmly in the past, traditional knowledge (TK) is both cumulative and dynamic, building upon the experience of earlier generations and adapting to the new technological and socio-economic changes of the present (Johnson 1992). Compared to

Geertz’s emphasis, TEK highlights the applied role of such a knowledge system and is little interested in symbolic or other theoretical aspects.

The concept is very popular with activists and applied researchers in all fields, which involve the local and sustainable development of resources, the critique of outside involvement in such developments and most notably the implications of the Convention Chapter 1 Introduction 25 on Biological Diversity (CBD or Convention of Rio de Janeiro, 1992) (Secretariat to the

Convention on Biological Diversity 2001). Despite its at first glance apparent limitations to ecological knowledge, it normally encompasses botanical, zoological geological, pharmaceutical and many other aspects of local knowledge. Many people now recognise the enormous contribution that TK systems have made and can make in the future both to the conservation and the sustainable use of biological diversity. Most indigenous and local communities live in regions where the vast majority of the world's plant genetic resources are found. Simultaneously these activists and scholars argue for urgent action to safeguard such TK and for a broad view of how humans and the environment interact.

Importantly, this discussion proceeded with a focus on non-European indigenous groups while far less attention has been paid to the situation within Europe.

The origin of TEK is based on the knowledge that societies existed under conditions of constant pressure on the resources upon which they depended, and that a means had to be found to persuade communities and families to economise with regard to their use of natural resources (Anderson 1996). TEK is a constantly evolving way of thinking about the world. Although views covered by TEK are described as “traditional”, this does not mean that they do not change or that it would be desirable that they do not adapt to new needs. Each generation makes their own observations, and compares their experiences with what they have been taught (Pierotti & Wildcat 2000). Slikkerveer (1999) described this knowledge system as “not just ‘simple’, ‘static’ or ‘archaic’, but rather dynamic with elements of both continuity and change, embedded in their adaptive capacity, selection mechanisms and appropriate use”. The practice of TEK differs from that of scientific ecological knowledge in that it is largely dependent on local social Chapter 1 Introduction 26 mechanisms (Berkes et a l 2000). Social mechanisms often play a role in the integration of ecological knowledge of different kinds. Mechanisms for the intergenerational transmission of knowledge are embedded in social systems. Successful transmission of agricultural skills and knowledge, for example, depends on the amount of time families spend on the land because of apprenticeship-based knowledge transmission, and the amount of time required for hands-on learning.

These are examples from a diversity of human activities as they relate to humans’ use of the environment. The next section highlights the link specifically as it relates to the use of local food in the Mediterranean basin.

1.2 Local foods of the Mediterranean

Food is an example which exemplifies local knowledge or TEK. Plants, fungi and animals that yield food are one of the most essential resources for all humans and of course an everyday need. At the same time, in many regions food is still produced or gathered in the immediate surroundings of the house or the community (Heywood

1999b). It is a very basic need, but - provided people do not suffer from starvation - also a pleasant experience. Food and dishes do always reflect a “vision of the world” and consequently peoples, ethnic groups and communities are proud of their special dishes and the plants or breeds of animal they produce and use. Such knowledge gives them a regional identity. However, it is well beyond the scope of this introduction to discuss the concept of “Local Food” on a world-wide scale. Chapter 1 Introduction 27

There are still many rural regions and communities in Southern Europe which follow traditional life styles. This includes the use of vegetables and fruits gathered in the wild.

Southern Europe (Greece, Albania, Italy, France, Spain, Portugal) harbours a large number of botanical species, a great proportion of which are endemic (Eberle 1975). The

Mediterranean TFK has sometimes been described as a diet mainly composed of pasta, olive oil, vegetables, fruits, red wine, seafoods and only a little red meat and was termed the “Mediterranean diet”. More precisely Mediterranean TFK should be referred to as

“Mediterranean diets'’'' since the many different cultures, religious beliefs, écologie backgrounds and historic developments around the Mediterranean basin resulted in many diets, which share a multitude o f elements, but also revolve around distinct local or regional traditions.

How can “local food” be defined? In a geographical approximation, this clearly relates to ingredients which are grown or produced locally by individuals or gathered, for example by shepherds, and prepared into dishes which often represent a local speciality.

Such food is derived from animals, fungi and plants, but in the context of this discussion the focus is on food of botanical origin. Some of these local traditions have become world-wide phenomena (in a way a “global food”) but this binary opposition does not really help in explaining the differences between what is only known locally with more widespread types of food. If one were to create a binary opposition, the alternative to local food would be food, which is commercialised well beyond the region where it is produced. Chapter 1 Introduction 28

Globalisation has resulted in an increasing availability of similar foods in different cultures. Local knowledge from Naples, Italy for example was transformed into a worldwide food, now often considered to be a typical American dish. The pizza was brought over to the USA and on its journey back to many European countries it became an element of US-American cultures (even though without the discovery of the

Americas the Aztecan “tomatl” would not have found its way onto the Napolitanean pizza) and is now a prime example of a convenience food.

This incorporation of new food species is, of course, not a new phenomenon, Turkish com (Fuchs’ Türkisch Kom) is included in many of the early European herbals like

Fuchs (1501-1566) as Turcicum frumentum. In fact, it refers to Zea mays, which, of course, is of Mesoamerican origin (Finan 1948). Similarly, sweet and hot peppers

{Capsicum annuum and C. frutescens) are now considered typical elements of the

Hungarian and Balkan (as well as the Asian Indian) cuisine, but are again of American origin. A telling example comes from the Mediterranean. In recent years rocket salad

{Eruca sativa, Brassicaceae) has become a new, popular food in certain sectors of the

European Community. Some health benefits have been scientifically investigated

(Hocking 1997; Esiyok et a l 2004; Barillari et al 2005). The aerial parts were originally well known only in the Central provinces of Italy (esp. Umbria) and it was an important food of the Ancient Romans, that later fell into disuse in most regions (Padulosi &

Pignone 1997). Remnants of this widespread reputation in ancient times are its persisting local use as a flavouring agent in traditional dishes such as gazpacho of North Murcia and East Albacete provinces in Southern Spain (Rivera & Obon 1991). Its leaves have a prominent, spicy taste and in recent years have become common ingredients of many Chapter I Introduction 29 salads in North and Central European countries. Thus rocket salad is a prime example of the transformation of a local food into a commercial product. Thus local food may well have its origin outside of the regions of use and the crucial aspect of its definition is the local production, gathering or harvesting and consumption.

As exemplified above, history shapes food cultures (Mennell et al. 1992). Another core element of food culture is the biodiversity and availability: traditional cuisines are obviously very dependent on the natural resources available in the area. Due to the importance of local geographical conditions for the availability of food items, regional patterns have traditionally been particularly relevant for food products. Such regional patterns persist today even if modem distribution systems have liberated local eating patterns from the constraints of climate. This is also exemplified in the slow changes of food habits in immigrant societies (Mennell et al 1992; cf. Nebel 2005). Wild gathered food is often only available seasonally and is consumed either fresh (e.g. spring salads and vegetables, fruits in autumn) or in a conserved form (dried, fermented, pickled).

Gathered food plants, often weedy, synanthropic or wild species grow in primary or secondary habitats. Some are strictly endemics others are widespread species (Inocencio et al. 2003).

Socio-cultural factors and history determine the roles of wild gathered food plants and their importance is largely based on taste, palatability, toxicity, cultural taboos and difficulties in production or collection. Some less appreciated local food plants are consumed only as famine food. These species have particularly been used in times of war or during seasonal chronic shortages (Tardio et al. 2006). They are not consumed Chapter 1 Introduction 3 0 regularly due to factors such as accessibility, or palatability. Often these foods are culturally associated with marginalised classes or peoples from culturally and economically deprived areas.

1.3 The changing food pattern

Food use is changing very fast on all continents (Haddad 2003; Popkin 2004). The dietary transition is driven by changing preferences fuelled by growing incomes, changing relative prices, urbanisation, populations growth, climate change and food technology. In industrialised countries, there is a considerable rise in expenditure for convenience food (pre-prepared or ready made dishes). At the same time, nutraceuticals and functional foods are a rapidly growing segment of the market (Arvanitoyannis &

Van Houwelingen-Koukaliaroglou 2005). The latter is specifically in response to risk factors that impact on the health of everyone. A series of risks are highlighted in the numerous studies focusing on diet and health (Willett 1994; Hu 2003; Biesalski 2004;

Bes-Rastrollo et al 2006; Panagiotakos et al 2006), including:

• Overconsumption of energy (calories)

• High intake of glucose

• Low intake of fruits and vegetables (bioactive compounds)

• Low intake of fibre

• High intake of red meat

• An imbalance in the ratio of omega 3:6 fatty acids Chapter 1 Introduction 3 1

Aside from dietary aspects other problems, such as environmental factors and lack of activity have been implicated (Wannamethee & Shaper 2002; Kennedy 2006). This increases the risk of numerous diseases including various forms of cancer (Donaldson

2004), cardiovascular (e.g. Parikh et al. 2005) and skeleto-muscular diseases, but also numerous metabolic syndromes like diabetes. Taking diabetes as an example, it is now apparent that not only people in many developed countries suffer from the various forms of diabetes and the illnesses associated with it, but in the last few decades the disease has also become a serious health problem in indigenous and local communities all over the world, including Mexico, the USA, Canada, Australia, but also in many regions of Asia and Polynesia (Fujimoto 1996; Shaw & Chrisholm 2003; Rivera et al. 2004; Andrade-

Cetto & Heinrich 2005; Ramachandran 2005). Clearly, there is a direct link between changes in dietary traditions and the overall consumption of the various classes of foods and the increased prevalence of this syndrome. According to the World Health

Organization (WHO 2006), diabetes affects more than 176 million people worldwide and at least one in 20 deaths are attributable to diabetes. The changes in diet are difficult to evaluate since in many instances the “traditional” or local forms of eating have disappeared and normally no quantitative data on food consumption in traditional societies are available. For many policy makers it seems such knowledge only becomes a topic of research once it has disappeared and health and socio-cultural problems develop.

The Mediterranean region is an area with a diverse and relatively well known dietary tradition commonly called the “Mediterranean diet” (Keys & Keys 1975) and focuses specifically on local traditions which show a great degree of diversity. The nutritionist Chapter 1 Introduction 32

Ancel Keys found that firemen in Naples and Madrid had a significantly lower blood cholesterol level than Americans, and that this correlated with a lower percentage of fat in their daily diets. Keys and collaborators (Hoffman 1979; Keys 1980) also showed that major cultural differences exist in the distribution of risk characteristics and risk behaviours, including diet, and in the geographic and time distribution of the major causes of death. Keys demonstrated that traits, which previously had been considered irrevocable and constitutional, such as body type, blood fat levels (cholesterol), blood pressure, heart rate, and responses to stress, were, in fact, largely modifiable by simple changes in the composition and quantity of diet and physical activity and that a

“Traditional Mediterranean Diet” helps to improve health parameters (Keys & Keys

1975). More specifically, a clinical trial within the “Lyon Heart Study” concluded that alpha-linolenic acid-rich diet was able to lower the rate of cardiovascular infarction (de

Lorgeril et a l 1999). Sources of omega-3 fatty acids like alpha linolenic acid are green vegetables, nuts, seeds, game and wild seafood, while wild gathered vegetables are especially rich in alpha-linolenic acids and antioxidants (Lionis et a l 1998; Crawford et a l 2000; Zeghichi et a l 2003; Biesalski 2004).

1.4 Food as medicine - medicine as food

Food is far more than nutrition or culinary art. The Ancient Greeks believed that a proper diet is a prerequisite to physical and intellectual well-being. It is now generally accepted that in traditional societies plants may be used both as medicine and food. It is difficult to draw a separating line between these two categories, since they are cultural constructions: food may be medicine and medicine may be food (Etkin & Ross 1982;

Etkin 1996; Heinrich 1998; Johns 1999; Pieroni & Price 2006). Chapter 1 Introduction 3 3

Many wild plants have multiple uses, for example providing nutrients, adding to dietary diversity, or providing medication for humans or livestock. In human nutrition, it is important to consider the overall benefit from such multiple use, taking into consideration not only the direct nutrient contribution from wild edible plants, but also the potential contributions of “medicinal foods” (Etkin & Ross 1982, 1994; Craig 1997;

Gibson 1998; Milner 1999; Gurib-Fakim 2006).

Interest in healthy nutrition has increased considerably in recent years, underlining the importance of this food-medicine interface. Today, more attention is being paid to positive or preventive nutrition due to the increased choice of food products with the desired functional components. In the USA, Europe and Australia the “non-nutritive” health roles of vegetable and fruit consumption are receiving increasing attention within the areas of functional foods, nutraceuticals, or phyto-nutrients (Balentine, Albano, and

Muraleedharan 1999; Craig 1999; Lampe 1999; McNutt 1995; Wahlqvist 1992). Current research on the disease-preventing roles of food plants has revealed a number of bioactive substances with potential disease-preventing or health promoting mechanisms

(Craig 1999; Lampe 1999), including components which have antioxidant activity, stimulate the immune system or have antibacterial or antiviral activities (Rice-Evans &

Packer 2003; Halliwell et al 2005; Riso et a l 2006).

Plant biodiversity is essential to human health, because plants are a source of both nutrients and medicinal agents and they contribute to socio-cultural well-being.

Conservation of biodiversity and the knowledge of its use are necessary resources for present and future health. The following model by Johns (Figure 1) recapitulates the Chapter 1 Introduction 34 relationship between biodiversity, diet and health in the context of contemporary global environmental and socio-economic change (Johns 2001):

IndustrialisationPopulation IndustrialisationPopulation growth ENVIRONMENTAL DISTURBANCE Urbanisation Climate change Biodiversity loss

Optimisation DIETARY / FUNCTIONAL DIVERSITY BIOCULTURAL Inadequacy DIVERSITY

Oxidative Malnutrition HEALTH status CHALLENGES Toxicants & Immunity & Xenobiotics Infection

Figure 1: Model of Biodiversity, Diet and Health in the context of global change (adapted from Johns 2001)

1.5 The “Local Food-Nutraceuticals” project and its goals

The local traditions of the Mediterranean were the focus of the project “Local Food-

Nutraceuticals” (LFN), in which this PhD project was embedded. The LFN project included specialists in many fields like ethnobotany, pharmacognosy, pharmacology and nutritional studies, which came from the UK, Germany, Poland, Spain, Italy,

Switzerland and Greece (list of LFN members see Appendix A in The Local Food-

Nutraceuticals Consortium 2005). This project had a very strong focus on ethnobotanical Chapter 1 Introduction 35 questions and was truly multidisciplinary bridging the gap between socio-cultural and natural sciences. The interest in local foods derived only in part from its potential as a lead source for novel nutraceuticals, and mainly focused on local food as an important element of local identity and as a link of a community with its history.

On the one hand the Consortium contributed to the development of new nutraceuticals by identifying plants traditionally used in rural communities of Southern Italy, Greece and Southern Spain with potential health beneficial effects, but as importantly the project gave new values to local food products which have been used for many generations and which now are on the brink of becoming forgotten.

Specific goals of the project included:

1. Ethnobotanical documentation of food products of selected communities in

Southern Italy, Spain, Greece inch Crete

2. Comprehensive understanding of the social, cultural, economical framework of

local food use

3. Identification of active extracts/pure compounds (leads for new health food

supplements)

4. Biochemical/pharmacological in vitro mechanisms/ in vivo effects of selected

species

5. Dissemination of ethnobotanical information in local/national languages.

There were two distinct but connected lines of research activities in this project - the study of local food knowledge with the goal of making this of interest to future Chapter 1 Introduction 3 6 generations in the regions of origin and thus to contribute to sustainable local use of resources, and the study of the species’ wider potential as nutraceuticals/health foods.

These two lines were in no way based on contradictory goals, but in fact interdependent aspects of one common goal - the valorisation of local knowledge. In order to ascertain appropriate benefit sharing between the regions of origin and the academic institutions involved in the project, the Consortium had given itself a precise framework in the form of a Consortium agreement (for details see Heinrich et al. 2005b). Chapter 2 Ethnobotany 37

2 Wild food plants used in the Graecanic area in Calabria, Southern Italy

Nino with wild gathered fennel {Foeniculum sp.)

Galliciano 2002 Chapter 2 Ethnobotany 38

2.1 Introduction

The Mediterranean basin has a long and multifaceted cultural history and harbours a high biodiversity (Eberle 1975; Hammer et al 1987; Medail & Quezel 1997, 1999;

Lardos 2006), housing approximately 25’000 species, about half of which are endemic to the region (Heywood & Skoula 1999). In general, biodiversity has provided the basis for the development of many agricultural crops and the Mediterranean area belongs to one of the most important centres of cultivated plant origin and diversity (Harlan 1971;

Kurlovich et al 2000). Thus, biodiversity is strongly linked with food, nutrition and human health. All of the world’s major food and fibre crops are of ancient origin and have had an anthropogenic evolution (Harlan & de Wet 1973). Local knowledge on plants and biodiversity are linked intrinsically, as human’s activities heavily impact on the components and attributes of biodiversity (Gadgil et a l 1993; Prance 2000). From both a botanical and anthropological perspective minor field crops or ones derived from managed non-cultivated species are of particular relevance and specifically in attempts to protect a region’s biodiversity.

Since antiquity humans have used a great diversity of non-cultivated plants, also commonly referred to as “wild”^ plants in botanical literature (Heywood & Skoula 1999;

Tardio et a l 2006) for food, medicines, fuel and many other purposes (Harlan 1975;

Heywood 1999b). Today, wild food plants (WFPs) still play a substantial role in the

* In this study, the term “wild food plants” is defined as species/plants that grow in natural or semi-natural ecosystems and that can exist independently of direct human action. The term is contrasted with plants that have arisen through human action, such as selection or breeding and depend upon management for their continued existence. Chapter 2 Ethnobotany 39 traditional livelihood of rural Mediterranean areas (Heinrich et al. 2005b; Leonti et al

2006), although the traditional food knowledge has already disappeared from many regions or is currently at risk of becoming forgotten. In the past decade, only a few studies have systematically analysed the consumption of non-cultivated botanicals in the

Mediterranean area (Forbes 1976; Paoletti et al 1995; Bonet & Valles 2002; Guarrera

2003; Pieroni et a l 2005; Rivera et a l 2005; Tardio et a l 2005). All these studies clearly demonstrate that these local foods still represent a relevant part of traditional

Mediterranean diets.

Until very recently this farmers’ or shepherds’ knowledge has been transmitted only orally. So far, there have been only few attempts to understand the historical development of food or medicinal plant use in cultures in which knowledge is predominately transmitted orally (e.g. Leonti et al 2003b; Fiore et al. 2005; Heinrich et a l 2005a; Kufer et a l 2005). Establishing the historical depth of food and medicinal plant use is relevant from a variety of perspectives. Such an analysis allows understanding the historical development of plant use in a discipline largely relying on diachronic methods, and, as importantly, the diverse and complex relationship between humans and their environments (Balée 1998, p. 2) specifically as it relates to non- cultivated food plants. Knowledge about food plants and their use in the past, especially about frequent species, may be obtained with the help of methods from different fields, such as archaeobotany or historical disciplines (Waterlow 1989; Wilkins et al. 1996;

Pennington & Weber 2004; Bouby & Billaud 2005; Megaloudi 2005). Historical linguistics (also comparative linguistics), which is primarily the study of ways in which Chapter 2 Ethnobotany 40 languages change over time, might proof to be a useful tool in this context (Blench

2003).

In this chapter plants traditionally consumed in rural communities in Southern Italy are described from an ethnobotanical as well as historical-linguistic perspective. Methods from both fields were used for analysing ethnobotanical data from the Graecanic area in

Calabria, Southern Italy, in order to better understand the historical development of using local biodiversity in a traditional, oral knowledge system, and, more specifically, to understand the ethnobotanical significance of local food use from an anthropological and linguistic-historical perspective.

2.1.1 The ethnographic background: Graecanic communities in Calabria, SI

The migration of the Greek and other peoples in the Mediterranean in history is reflected in many ethnic, linguistic and religious minorities which still exist today (Rother 1989).

This study focuses on relatively isolated Greek communities in Calabria, Southern Italy.

Historically, parts of Southern Italy, as we know it today, came under Greek influence during the 8* century BC, and were known as Magna Graecia. The Greek influence continued over centuries until the end of the Byzantine Empire, also called the Eastern

Roman Empire (Cerchiai et a l 2004). Today, the Greek minorities in Southern Italy

(Graecanic areas) have receded into the eastern Province of Salento, Region of Apulia, and into the Province of Reggio di Calabria in the Region of Calabria (see Figure 2)

(Rother 1989; Pan & Pfeil 2000, p. 89-92). Chapter 2 Ethnobotany 41

ROME

K

CondoW Mef Manna di Pto. Salvo

Figure 2: Map of Italy. A - Region of Figure 3: Graecanic area in the Province of Apulia; S - Province of Salento; C - Reggio di Calabria, Region of Calabria. Region of Calabria; RC - Province of Graecanic villages: Amendolea, Bova, Reggio di Calabria Condofuri, Galliciano, Roccaforte del Greco and Roghudi (Condemi 1999)

The Graecanic communities in Calabria are located in the Aspromonte Mountains in the southern part of Apennine Mountains: Bova (in Grecanico: Viia), Amendolea

{Amiddalia), Condofuri {Condochiiri), Galliciano (Gaddicianà), Roccaforte del Greco

{Vuni), and Roghudi (Richùdi) (see Figure 3) (Condemi 1999). The population of the villages varies from 100 to 1000 inhabitants. The inhabitants of the Graecanic area are, as compared to the surrounding Italian population, characterised by their own Greek dialect Grecanico, culture and history as an ethnic and linguistic minority. Chapter 2 Ethnobotany 42

Grecanico (synonym: Greco or Italiot-Greek), the “Greek of Calabria” belongs to the

Attic Greek subgroup of Greek (Indo European language family) (Gordon 2005). Two main theories about the origin of Grecanico, based on historical sources, as well as on historical linguistics (Sideras 1979), are discussed: the Byzantine theory and the continuity theory. According to the Byzantine theory the Greek-speaking population of

Southern Italy resulted from the immigration of different ethnic groups from Greece during the Byzantine Empire (330 AD - 1453 AD) (Comparetti 1866 [reprint 1976];

Morosi 1870 [reprint 1994]; Tozer 1889; Battisti & Alessio 1957). The continuity theory, on the other hand, argues for the continuation of Greek culture and language in this area since the colonies of the Magna Graecia (8* century BC) (Hatzidakis 1892;

Rohlfs 1964, 1972). Today, Grecanico is only spoken by elderly people, whilst the younger generation mainly communicates in the Calabrian dialect of Italian.

Furthermore, Italian influence has increased dramatically after the building of access roads to the villages in the 1950s.

The ethnic Greek minorities living in Southern Italy today exemplify the establishment of independent and permanent colonial settlements of Greeks in the past. With the foundation of settlements in Southern Italy, the Greeks took common crops like olives, or grapes with them to cultivate in the new colonies (Sallares 1991, p. 92; Lombardo

1996; Dalby & Brodersen 1998). It is a well-known phenomenon, as when people have travelled, traded and migrated, they have always moved their crops with them (Prance

2005). Today, pastoralism and subsistence agriculture are still the main traditional livelihood in the area (Figure 4). However, economic changes, emigration as well as serious floods and earthquakes (in 1951 and 1971) have led to the gradual abandoning of Chapter 2 Ethnobotany 43

tillage and pastoral activities and a sharp decline of the resident population. In addition,

extensions of the communities were built along the coast {la marina), and many

inhabitants left the mountain villages and moved to la marina (Kish 1953).

Nevertheless, the gathering of WFPs still plays an important role in the traditional diet

today and is an integral part of the Graecanic culture.

Figure 4: Shepherd with flock of sheep and goats, Galliciano 2002

2.1.2 Historical sources of botanical data

The wealth of written evidence that exists in Europe and other traditions (esp. Chinese,

and Ayurvedic) from Ancient times offers an important source of data for ethnobotanical

comparison. Written sources of Ancient Greek traditions include the botanical treatises

of Theophrastus of Eresos (372 - 287 BC, Classic) that have survived in translations and

represent a great source of botanical (including names, description and uses) and

horticultural information from antiquity (Historia Plantarum) (Hort 1916). Another very Chapter 2 Ethnobotany 44 important work is de materia medica from Pedanius Dioscorides (ca. 40 - 90 AD, Early

Roman) including therapeutic uses of plants (Gunther 1934). In all cases, botanieal identifications of the plants mentioned in these aneient texts are available (esp. Singer

1927; Tschirch et al. 1933; Steam 1976) and are mainly based on a.) description of plants; b.) information about habitats; c.) use and properties of described plants; d.) illustrations; and e.) vernacular names (phytonimes) of plants in use in Greece (Fortes

Fortes 1984; Baumann 1993, p. 11). The botanist de Toumefort (1656 - 1708) was able to identify a great number of plants of Dioseorides (Singer 1927). Many also were identified later by John Sibthorp (1758 - 1796 ), who travelled extensively in the Eastern

MediteiTanean (Singer 1927). However, the botanical identification of Ancient Greek plants mentioned by authors like Theophrastus or Dioscorides can only be ascertained with a certain degree of ambiguity (see Figure 5 as an example of a drawing).

3H DIOSCORIDES, BK. Ill Figure 5; Drawing of

Marathron - Fennel -

Foeniculum vulgare

(Gunther 1934)

MAHATflllON

8 i. Matiathron . Foeniculum vulgare Fennel Chapter 2 Ethnobotany 45

As Grecanico is only a spoken language, and there is no written evidence in Grecanico available from the past. The linguist Gerhard Rohlfs listed some Grecanico plant names in his Lexicon Graecanicum Italiae Inférions (1964) and analysed the etymology from a linguistic point of view. However, his plant identifications were only very vague, often misinterpreted and rarely associated to a specific use.

2.2 Aims and Objectives

The principal aim of this PhD project was investigate the current role of WFPs in the traditional diet in the Graecanic communities in Calabria, Southern Italy and their potential health properties. The objectives were to:

• identify WFPs traditionally consumed in the Graecanic area

• understand the historical importance of wild gathered food plant species

• examine local beliefs and multifunctionality of WFPs

Research Questions

What WFPs are gathered in the Graecanic area? Where and when are they gathered?

Who gathers them? How often are certain species gathered? How are they prepared and consumed? How do local people perceive the taste of these WFPs? Do local people consider WFPs to have any health beneficial effects? Are these plants used for other purposes (multifunctionality) than food? Chapter 2 Ethnobotany 46

2.3 Methods

Ethnobotanical fieldwork was mainly conducted in Galliciano (Community of

Condofuri, see Figure 6) during the spring season of 2002 and 2003 and two months during the autumn of 2002. Additional fieldwork was carried out in the other Graecanic communities of Bova, Ghorio di Roccaforte, Roghudi, Amendolea and Condofuri in

2003. Permits for research, collecting and export of voucher specimen were obtained from the authorities of the Aspromonte National Park (Direzione Ente Parco Nazionale delP Aspromonte) and the community of Condofuri. The identification of the food plant species is based on Pignatti’s Flora d ’Italia (2002). Two voucher specimens of each food plant accompanied by detailed notes on the collection locality, characteristics of the plant, its local culinary uses, vernacular names and their meanings were deposited at the herbarium of the Centre for Pharmacognosy and Phytotherapy, School of Pharmacy,

University of London (UK). Chapter 2 Ethnobotany 47

Figure 6: Galliciano (municipality of Condofuri, RC), in the southern part of the Aspromonte mountain, 621 metres above sea level, approximately 120 inhabitants.

2.3.1 Ethnobotanical methods

Traditional knowledge regarding food plants was assessed using standard ethnobotanical tools, including free listing exercise (Alexiades & Sheldon 1996, p. 53—94; Quinlan

2005), participant observation (Martin 1995, p. 96) and interview techniques (Schensul et al. 1999). In the first phase of the field research 18 research participants (7 male and

11 female; aged between 18 and 88, average age 55) were asked to list any non- cultivated food plant that are used on a regular basis or were used in the past respectively, to obtain information about the cultural domain of WFPs. Data obtained from the free listing exercise were analysed using ANTHROPAC (4.72 version)

(Borgatti 1992). Chapter 2 Ethnobotany 48

M i

Figure 7: Signora Tota and the author, Sabine Nebel, during an interview.

In-depth knowledge about the use of the identified WFPs was obtained through semi­ structured and structured interviews (Schensul et al. 1999) with 36 elderly inhabitants of the Graecanic communities (average 64 years old; 19 female, 17 male) (see Appendix

8.1 for ethnobotanical questionnaire schedule). The interviews (e.g. Figure 7) were all conducted in Italian, as all informants in the Graecanic communities are bilingual in

Calabrian (an Italian dialect) and Grecanico. For each of the identified food species the local name in Grecanico, if available, information about the plant parts used, the traditional culinary uses, taste and frequency of use were recorded.

Participant observation techniques (Martin 1995; Cotton 1996; Cunningham 2001) were utilised to better understand the cultural implications of plant gathering, preparation and Chapter 2 Ethnobotany 49 distribution of foods within the community. Participant observation proved to be a useful method for discovering details about social and ecological issues. Classical cognitive anthropological series of queries were also used to analyse the local classification of these botanicals (Berlin 1992).

2.3.2 Historical-linguistic method

To better understand the cultural and historical importance of wild food plant species in the Graecanic area, vernacular plant names of edible plants were compared with plant names mentioned in Ancient Greek literature (Fraas 1845; Lenz 1859 [reprint 1966];

Langkavel 1866 [reprint 1964]; Hort 1916; Bois 1927; Gunther 1934; Steam 1976;

Baumann 1993; Raven 2000; Hondelmann 2002) and vernacular names in Modem

Greek literature (Heldreich 1862; Candargy 1889; Fragkaki 1969; Yannitsaros 1977;

Tzanoudakis 1982; Lambraki 2000; Sawides 2000; Lardos 2006). If one compares two geographically distant but linguistically related groups, cognates give a good indication of the historical importance of locally named species (Leonti et al 2003b). Cognates are, in historical linguistics, pairs/sets of words in related languages that have developed from the same ancestral word (Trask 1996, p. 205). However, cognates of plant names have to be treated with caution, since not always has one scientific plant species one vemacular name {one-to-one-correspondence), but sometimes one plant species might have more than one folk name {over-differentiation) or two or more scientific species might have the same vemacular name {under-differentiation) (Berlin 1973). Chapter 2 Ethnobotany 50

2.4 Results and Discussion

2.4.1 Wild food plants traditionally consumed in the Graecanic area

The inhabitants of the Graecanic communities in which the study was conducted regularly gather 48 wild food plant species, including wild greens, fruits, condiments and some mushrooms. Table 1 lists food plants cited by at least three informants

(cf. Johns et al 1990) and includes botanical and ethnobotanical core information on these plants.

The plant family contributing most members to wild gathered food plants are the

Asteraceae with a total of 20 species (42 %). The second most important families are the

Apiaceae, Brassicaceae, and Rosaceae with three species each. Table 1: WFPs traditionally gathered and consumed in the Graecanic communities in Calabria (° Galliciano, Roccaforte del Greco, "^Amendolea, ^Condofuri, ® Bova, Roghudi, only indicated, if the plant name is specific for one village); FU = Food use: r = raw; nr = non raw; b = both; Taste = pricio (bitter), glicio (sweet) FRQ = Frequency of use: ••• = once a week; •• = less than once a week, at least once a month; • = less than once a month

Vernacular name Scientific plant name Family Part(s) used FU Traditional culinary uses Taste FRQ {Grecanicd)

Apium nodijlorum (L.) Lag. spèlendro (singular), Apiaceae aerial parts b raw or cooked as salad, with spring celery e spèlendra (plural.) onions, oil and vinegar like Asparagus acutifolius L. asparagi Liliaceae young shoots nr Frittata = omelette with egg, flour glicio * and pecarina (goat’s cheese) Boletus edulis Bull. Fr. bavoselli, bavuso, Boletaceae fruiting body nr - boiled with rice or pasta bavuseddhi - with tomato sauce

Baraga officinalis L. gabbuina*^ Boraginaceae aerial parts b Chorta tiganimena (CT)‘ and glicio # vramena (CV)“ ligabbuina* Capsella bursa-pastaris (L.) - Brassicaceae young leaves nr CT,CV# Medik. Carlina carymbasa L. fucida Asteraceae stem r raw as snack glicio #

Castanea sativa Mill. castano, castana Fagaceae fruits b - raw as snack = castana omà ••• - baked = castana furrimena - boiled in water = castana vramena Chandrilla juncea L. grospartu Asteraceae young shoots, basal b - raw as snack (peeled and eaten) ## leaves - as salad with oil and vinegar - CT,CV

51 Table 1 continued

Scientific plant name Vernacular name Family Part(s) used FU Traditional culinary uses Taste FRQ Chrysanthemum segetum L. cucuddho, Asteraceae top of aerial part r raw as snack pungent • cucuddhaci Cichorium intybus L. cicoria Asteraceae basal leaves nr CT, CV pricio #«

Cynara cardunculus L. ssp. carciofi salvacce Asteraceae flower bud nr boiled and/or fried • cardunculus

Ficus carica L. SI CO Moraceae pseudofruits - eaten fresh as snack (diverse cultivars) - dried as snack, eaten in winter = Cozsula - Petràlia = traditional sweet prepared during Christmas period with a mixture of honey (meli), flour, almonds (amiddala), walnuts (caridi), dried figs and raisins. Foeniculum vulgare ssp. masaro (sg.), masara Apiaceae young aerial parts b - Fasuli me ta masara: soup with glicio piperitum (Ucria) Coutinho (pi.) fenocchio and seeds beans, wild fennel, potato and pasta salvacce ® - as condiment in salads - to spiro masaro = fennel seeds used as condiments in local produced salami and other dishes - CT, CV

52 Table 1 continued Scientific plant name Vernacular name Family Part(s) used FU Traditional culinary uses Taste FRQ Hedypnois cretica Willd. - Asteraceae basal leaves nr CT, CV glicio ###

Hedysarum coronarium L. sùddha Fabaceae stems r raw as snack glicio #

Hirschfeldia incana (L.) Lagr.- senapia Brassicaceae aerial parts nr CT,CV e# Foss. Hypochoeris achyrophorus L. maruddhaci, Asteraceae basal leaves nr CT, CV glicio •• maruddhi Hypochoeris radicata L. costardeddhe, costa Asteraceae basal leaves nr CT,CV ••• di vecchia Juglans regia L. caridi (walnut) Juglandaceae kernel b - raw as snack ••• caridia (walnut tree) - in sweets like Petràlia (see Ficus carica) Lactarius deliciosus (L.) Gray rusitte Russulaceae fruiting body nr first boiled, then fried with olive oil, • farina and garlic Lactuca serriola L. glossa to sciddhio Asteraceae young aerial parts b CT,CV glicio •

Lactuca viminea (L.) J. Presl pricaddhlda Asteraceae basal leaves b - raw as salad (with oil and vinegar) pricio ••• & C. Presl - CT,CV Laurus nobilis L. lauro Lauraceae leaves nr as condiment in the traditional ••• tomato sauce

Lotus edulis L. carrafucia® Fabaceae young fruits r raw as snack glicio • carrambatula*

53 Table 1 continued Scientific plant name Vernacular name Family Part(s) used FU Traditional culinary uses Taste FRQ Mentha sp. menta Lamiaceae aerial parts nr Peperonata = sweet pepper, potatoes •• and aubergine fried in olive oil, condiments: mint and rosemary Moms alba L. sicaméno aspro Moraceae fruits r raw as snack •

Moms nigra L. sicaméno mavro Moraceae fruits r raw as snack •

Onopordum illyricum L. anapordo ° Asteraceae inner part of roots r raw as snack glicio •

napordo'^ Opuntia ficus-indica Mill. sico tu trucu, Cactaceae fruits r - raw as snack ••• ficarazzi (fruits); - ta ascasia (ficarazzi afflirimena) pittara (plant) dried fruits eaten as snack during winter Origanum heracleoticum L. rigano Lamiaceae flowering tops b condiment •••

Papaver rhoeas L. paparina Papaveraceae basal leaves; leaves nr CT,CV glicio •••

Pimpinella anisum L. cinimo Apiaceae seeds nr Scardatèglia: aniseed flavoured • sweets prepared for weddings

Portulaca oleracea L. andracla°, Portulacaceae aerial parts r raw as salad (oil and vinegar) glicio • porcellana^

54 Table 1 continued Scientific plant name Vernacular name Family Part(s) used FU Traditional culinary uses Taste FRQ Prunus dulcis (Mill.) D. A. amiddala Rosaceae kernel r Petràlia (see Ficus carica) ••• Webb. mandorlata = almond milk Punica granatum L. rùdi Punicaceae fruits r raw as snack •••

Reichardia picroides (L.) gaddhazzida® Asteraceae basal leaves b - raw as snack glicio ••• Roth. caddhazzida^° - CT, CV galazzida® - Gaddhazzida me ta fasuli (with beans) Reseda alba L. gattinaria ^ Resedaceae aerial part b - Eaten raw with olive oil and bred pricio ••• - CT, CV Rosa canina L. rosa Rosaceae petals r - Meli (honey) flavoured with rose • petals (dried) - Ri cotta eaten with rose-honey Rubus ulmifolius Schott mure che chalipo Rosaceae fruits r raw as snack ••

Rumex acetosa L. aceturia Polygonaceae stems r raw as snack pricio •

Rumex scutatus L. aceturia Polygonaceae leaves r raw as snack pricio • Scolymus hispanicus L. sculimbri Asteraceae leaf stalks nr - Sculimbri me ta lasagne: leaf stalks glicio •• cooked with home made pasta - CT, CV

Silene vulgaris (Moench) cavuraci Caryophyllaceae young aerial parts nr CT, CV glicio • Garcke cavuleddhu^

55 Table 1 continued Scientific plant name Vernacular name Family Part(s) used FU Traditional culinary uses Taste FRQ Sinapis arvensis L. rapa Brassicaceae young aerial parts nr Frittata = omelette with egg, flour •• and pecorino (goat’s cheese) Sonchus asper (L.) Hill. zuccho Asteraceae young leaves nr CT,CV glicio ••

Sonchus oleraceus L. zuccho Asteraceae basal leaves nr CT,CV glicio •••

Taraxacum sp. cuscuneddhi, Asteraceae basal leaves nr CT,CV pricio ••• cicoria Tragopogon crocifolius L. Asteraceae young leaves b - raw as snack glicio • - CT,CV Tuber melanosporum caratompola Tuberaceae fruiting body b - raw •• Vittadini (symbiotic with - grated on top of pasta Cistus sp.) - cooked, sliced and fried with farina in olive oil - sotto olio - preserve in olive oil

Urospermum dalechampii russéddha^ Asteraceae basal leaves b - raw as snack • (L.) Schmidt - B. - CT, CV Urospermum picroides (L.) zuccho Asteraceae basal leaves nr CT, CV pricio •• Schmidt

‘ Chorta tiganimena (CT) = fried wild edible greens {chàrta) washed then boiled and fried in olive oil with garlic and chili; “ Chàrta vramena (CV) = cooked wild edible greens {chorta) boiled in salted water until tender. Eaten with olive oil and lemon.

56 Chapter 2 Ethnobotany 57

Elderly women are the main keepers of traditional knowledge in the domain of local food plants (Figure 8). Women are directly involved in work with food for the family and therefore possess knowledge especially as it relates to different aspects of food production and use. Men most notably retain specific knowledge about gathering (e.g. identification, where, when) of WFPs. They play an important role in gathering plants and fungi that grow far away form the village. Only very few of the younger generation are able to identify the culturally most important wild edible plant species.

Figure 8: Signora Angela, gathering sculimbri - Leaf stalks of basal leaves of Scolymus hispanicus are prepared to be cooked with home made pasta {Sculimbri me ta lasagne) or together with other wild greens. Chapter 2 Ethnobotany 58

The gathering of wild greens is seasonal. Most of the traditionally used wild species are gathered during winter and spring (December - May) and they represent an important part of the daily diet. Historically, winter is considered the worst time of the year because of the limited range of foods available. It is during this period that the wild gathered vegetables offer dietary diversity (Forbes 1976). They are very valuable nutritionally by supplying vitamins, minerals or antioxidants, at a time when garden vegetables are not yet ready to be harvested.

Most of these WFPs are collected by women in vineyards {ambeli) or olive groves

(uliveto), which are normally located close to the village. A few species are also collected in hedgerows, or on fallow fields (see Figure 9). Generally, the men collect any wild species, including mushrooms, from the forested areas (bosco) on the Aspromonte

Mountain, which are located further away from the villages. Wild fruits like blackberries, or prickle pears are preferably gathered and eaten by children (Nebel 2005;

Tardio et al. 2005). Chapter 2 Ethnobotany 59

Ambeli (vineyards)

Figure 9: The surrounding area of Galliciano: Olive groves {uliveto), homegardens

{orto), vineyards {ambeli).

Ethnotaxonomically, edible greens are an intermediate category in the domain of food plants called ta chàrta in Grecanico, which roughly corresponds to what in bioscientific nutritional studies is called “green leafy vegetables” (Bender 2005; Pieroni et al. 2005).

Etymologically, ta chàrta has its origin in xoprog (Rohlfs 1964; Chantraine et al. 1968) and has two different meanings: “grassland”, but most of all “fodder, hay or herb”. In Chapter 2 Ethnobotany 60 modem literature, the same term la %6pra {ta chàrta or horta^) was found to describe

WFPs consumed in Greece (Forbes 1976; Lambraki 2000; Sawides 2000). This significant shift in meaning, from “animal fodder” in Ancient Greece to today’s “herbs for human consumption” (Chantraine et al 1968) is intriguing.

In general ta chàrta (see Figure 10) refers to non-cultivated species. However, because in many cases the distinction between wild forms of a certain species and the cultivated one is almost impossible, it is of limited relevance to the local people.

I

Figure 10: Bucket with wild gathered ta chàrta (wild greens)

“ transliteration of Greek characters into Latin characters according to ISO 843:1997 (http://transliteration.eki.ee/pdf/Greekpdf) Chapter 2 Ethnobotany 61

Other sub-categories in the domain of food plants (excluding fruits) are condiments, in

Grecanico àrtema and in Modem Greek bacharika (fXTiaxapiKd), or atrymata (aipupaxa) in Greek dialects (Lardos, personal communication). Etymologically, àrtema has its origin in ârtyma (apiupa) the Ancient Greek word for seasoning or condiment (Rohlfs

1964, p. 59). Interestingly, in the mid 19^ century the botanist Theodor von Heldreich

(1822 - 1902) also used the same popular term ârtyma (dpiupa) to describe wild greens or parts of greens, which are used as condiments in Greece (Heldreich 1862).

Mushrooms are another important food category, called mulitaria in the Graecanic area and manitària (pavndpia) in Modem Greek. The same term, manitària, was also used by Heldreich as popular name fox Agaricus camprestris L. and related species (1862).

In order to detect the culturally most salient plants of the domain “wild gathered food plants”, a free listing exercise was conducted (see Table 2). A relatively high salience was reported for pricaddhida {Lactuca viminea; 0.49, Figure 11), gattinaria {Reseda alba; 0.47, Figure 12) and gaddhazzida {Reichardiapicroides; 0.43, Figure 13). Chapter 2 Ethnobotany 62

Figure 11 : Young leaves of Pricaddhida {Lactuca t / - viminea, Asteraceae)

Figure 12: Inflorescence of gattinaria {Reseda alba, Resedaceae)

Figure 13: gaddhazzida {Reichardia picroides, Asteraceae) Chapter 2 Ethnobotany 63

Salience is a statistic accounting for rank and frequency and a high salience reflects both a high frequency and also high rank in the informants’ lists (these plants appear more often and earlier in freelists). Frequently mentioned plant species among individuals indicate common knowledge, or consensus, within the cultural domain. Accordingly, plants with a high salience are well known to both genders as well as in different generations. This is not the case for many WFPs: only very few of the younger generation, which are no longer fluent in Grecanico, are able to identify the culturally most important wild edible plant species. The consensus analysis, which enables the researcher to assess the degree of shared knowledge possessed by informants about a given cultural domain, showed a strong cultural agreement of 82 % (N =18). This high value indicates a high rate of agreement between the informants about the culturally most important wild food plant species.

Table 2: The most frequently mentioned wild food species in Galliciano, Graecanic area, Calabria, Southern Italy (free listing exercise with 18 informants (7 male and 11 female; Aged between 18 and 88, average Age 55). Analysed with ANTHROPAC, 4.72 version.

Vernacular Avg Scientific name Frequency* Salience Cognate name Rank Lactuca viminea pricaddhida 13 3.46 0.49 yes Foeniculum vulgare masaro 12 4.67 0.33 yes ssp. piperitum Reseda alba gattinaria 11 2.72 0.47 no Reichardia picroides gaddhazzida 10 3.00 0.43 yes Scolymus hispanicus sculimbri 8 7.50 0.10 yes Chapter 2 Ethnobotany 64

Table 2 continued

Vernacular Avg Scientific name Frequency* Salience Cognate name Rank Papaver rhoeas paparina 7 5.43 0.16 ** Sonchus oleraceus, or zuccho 7 6.29 0.16 yes S. asper, or Urospermum picroides Hypochoeris mamddhaci 7 4.42 0.23 yes achyrophorus * Frequency of vernacular name mentioned by the informants in free listing exercise. ** paparouna in Modem Greek; mékon roias in Ancient Greek (Gunther 1934). Etymology of plant name unclear, probably Latin or Italian origin.

Most of the gathered wild food plant species have vernacular plant names in Grecanico.

The correspondence between folk generic names and scientific names are mainly a one- to-one correspondence (Berlin et a l 1966). However, sometimes the same phytonym is used to name more than one similar botanical species {under-differentiation), as for example zuccho in Grecanico for Sonchus asper, S. oleraceus and Urospermum picroides. The same was reported in Greece, where the same three plant species are called tsochos, zochos, zôchos or agrio zochos (Heldreich 1862; Candargy 1889;

Lambraki 2000).

Of the total of 48 gathered species in Galliciano, more than half of the plant species

(58 %) today are also used as food plants in Greece (Lambraki 2000; Zeghichi et al

2003; Kypriotakis In prep.). As shown above, this does not come as a surprise, since these two regions have common cultural traditions, but also since the vegetation in the two regions is similar (Eberle 1975, p. 121). Most of the recorded plant species are also Chapter 2 Ethnobotany 65

used in other parts of Southern Italy and the Mediterranean, for example Papaver

rhoeas, Sonchus asper, or Reichardia picroides (Corsi & Pagni 1979; Aliotta 1987;

Paoletti et a l 1995; Bonet & Valles 2002; Pieroni et a l 2002b; Guarrera 2003; Zeghichi

et a l 2003; Mattirolo et a l 2005; Rivera et al 2005; Scherrer et a l 2005; Tardio et a l

2006). On the other hand, there are some species {Chrysanthemum segetum, Hedypnois

cretica, and Lotus edulis) that are predominantly used in Greece and in the Graecanic

area, but not in other parts of Italy (Picchi & Pieroni 2005). The use of these plants

might therefore be an element of Ancient Greek culture.

Reseda alba, gattinaria in Grecanico, is an example of a very local food plant, which is used very frequently and appreciated as vegetable in the Graecanic area, but not in

Italian communities, where the plant is considered to be too bitter in taste. In Galliciano,

the tops of the shoots are either eaten raw, seasoned with olive oil, or cooked and fried in

olive oil with garlic, chili and pepper mixed with other wild greens. In literature, only

two references to the use of Reseda alba as food in the Mediterranean were found.

Firstly, young leaves of Reseda alba used as vegetable in Greece (Heldreich 1862) and,

secondly, as salad by the villagers of the surroundings of Lamaca in Cyprus (Arnold

Apostolides 1991). Interestingly, both records are from regions of the Eastern

Mediterranean, which were, in historic times, part of the Greek and Byzantine worlds, as

the Graecanic area in Southern Italy.

Taste is a key criterion for perceiving, categorising and characterising food plants in

general (Grivetti 1981; Johns 1986; Nebel 2001). The inhabitants of the Graecanic area

often ascribe a specific taste to singular plant species, particularly to ta chàrta species Chapter 2 Ethnobotany 66

(see Table 1). For example the taste of Reseda alba, or Lactuca viminea is described as

“bitter” {pricio in Grecanico) and of Papaver rhoeas as “sweet” {glicio in Grecanico).

Local gatherers pay attention to collect both bitter and sweet herbs to assure a balanced taste of the dishes to be prepared. Generally, bitterness plays a very important role in the local perception of health, as the bitter taste of wild greens is perceived as healthy in the sense of “blood clearing” and “good for the liver”. In Roghudi, edible greens are also called chàrta pricia - bitter wild greens. Already in 1862 Heldreich reported that many plants from the Asteraceae family were considered to be healthy by the Greeks, because of their bitterness (Heldreich 1862). Pieroni et al. (2002b) reported similar results among ethnic Albanians in Southern Italy.

2.4.2 Traditional dishes made of wild food plants

Regional specialities and traditional dishes are important elements of local food culture.

Many dishes (see Table 3) made of green aerial parts or basal rosette of leaves are part of the everyday cuisine and consist of a broad range of species, reflecting the seasonal aspect of gathering, as well as the aspect of taste (bitter pricia vs sweet glicia). A traditional lunch in spring consists of pasta with tomato sauce (Italian sugo: tomato, onion, garlic, bay leaves, oregano or basil) as first course, and ta chorta (wild edible greens, see Figure 14), local bread, salad, salami and cheese {pecorino - sheep’s cheese)

as second course. Chapter 2 Ethnobotany 67

Figure 14: Chorta vramena - boiled wild greens.

Figure 15: Picture of “Fasùli me ta masara = Soup with broad beans, young leaves of wild fennel, carrots, potato, pasta and fresh chili. Chapter 2 Ethnobotany 68

Table 3: Examples of local dishes with WFPs in the Graecanic area in Calabria, Southern Italy

Name of the local Description, mode of Wild food plant species used dish in Grecanico preparation chorta vramena Mixed wild greens {chorta Reichardia picroides, Reseda (Figure 14) mimmend) boiled and alba, Lactuca viminea, Papaver seasoned with olive oil and rhoeas, Hypochoeris pipighe (chili) achyrophorus, H. radicata. Always; chorta pricia (bitter) Sonchus asper, S. oleraceus. mixed with chdrta glicia Urospermum dalechampii, U. (sweet) picroides chorta tiganimena Mixed wild greens {chdrta same species as chàrta vramena mimmena) boiled and sauteed in frying pan with olive oil, garlic and fresh chili fasùli me ta masara Soup with broad beans. Foeniculum vulgare ssp. (Figure 15) young leaves of wild fennel, piperitum carrots, potato, pasta and fresh chili sculimbri me ta Leaf stalks of Scolymus Scolymus hispanicus lasagne hispanicus cooked with home made pasta insalata di Raw or cooked as salad, with Apium nodijlorum spèlendra spring onions, oil and vinegar frittata Young shoots mixed with Hirschfeldia incana. Asparagus egg, flour and pecorino acutifolius, Sinapis arvensis (goat’s cheese) and fned like an omelette Chapter 2 Ethnobotany 69

The traditional preparations include boiling, frying, baking, or eating raw with olive oil and vinegar. Most leaf vegetables that need cooking can be added to soups, boiled and seasoned with olive oil, or sauteed in the frying pan with olive oil, garlic and other herbs.

2.4.3 Historical importance of wild food plants

The comparison of ethnobotanical data with historical sources (written evidence) in

Ancient Greek (e.g. Historia Plantarum from Theophrastus of Eresos, or de materia medica from Pedanius Dioscorides, and many more) suggests that people have used several WFPs since antiquity. The philosopher Plato, in his “Republic”, gives an example of a healthy diet, including olive oil, cereals, fruits and “country dishes of wild roots and vegetables” (Ferrari 2000, p. 55). This and other text passages permit to conclude, that already the Ancient Greek diet included a wide variety of wild greens, vegetables and fruits (Vickery 1936; Brothwell & Brothwell 1969; Fidanza 1979;

Sallares 1991; Lombardo 1996; Pearson 1997).

When comparing plants used as food in the Graecanic area with Greece, 15 cognates of plant species have been found (see Table 4). Interestingly, for all culturally important wild gathered food plants cognates have been reported, with Reseda alba as the only exception (see Table 2). Table 4: Cognates of chàrta (edible greens) and other WFPs used in Galliciano, Calabria, Southern Italy and in Greece

Vernacular names Graecanic area Vernacular names in Greece* Etymology of cognates Ancient Greek botany - possible (Italy) corresponding taxon, Greece according to Rohlfs (1964)^ identification^ corresponding taxon, plant family plant family asparagi sparaggi, sparagki asparagoi all kinds of tender shoots similar to Asparagus acutifolius Asparagus sp. Asparagus Liliaceae Liliaceae andracla andrakla, glystiridha andrachni Portulaca oleracea Portulaca oleracea Portulaca oleracea Portulacaceae Portulacaceae karidi (walnut) karidia (tree) karydi (walnut) karydia (tree) karydion small nut Juglans regia Juglans regia Juglandaceae Juglandaceae kromidi kremmÿdi krommy(d)ion onion {Allium cepa) Allium cepa L. Allium cepa L. Liliaceae Liliaceae gaddhazzida, galazzida galatsida, galaktida galactis (6th century AD) plant with milky juice (spurge, kind of Reichardia picroides Reichardia picroides thistle, or plant that is able to curdle milk); Asteraceae Asteraceae gala (gr.) = milk maruddhi, maruddhaci (both plural) agrio'* marulo, prikomarulo (singular) amarullion, marulion (6th kind of chicory? Hypochoeris achyrophorus and Lactuca serriola century AD) kind of lettuce? Taraxacum sp. Cichorium sp. Asteraceae Asteraceae

70 Table 4 continued Vernacular names Graecanic area Vernacular names in Greece^ Etymology of cognates Ancient Greek botany - possible (Italy) corresponding taxon, Greece according to Rohlfs (1964)^ identification^ corresponding taxon, plant family plant family masaro agriomarathos^, marathos marathron Foeniculum vulgare Foeniculum vulgare ssp. piperitum Foeniculum vulgare Apiaceae Apiaceae pricaddhida prikalida, pikralida pikralis, pikras kind of dandelion, bitter lettuce; bitter plant Lactuca viminea Cichorium sp. picros (gr.) = bitter Asteraceae Asteraceae rigano, riganaci rigani origanon Origanum sp. Origanum sp. Origanum sp. Lamiaceae Lamiaceae sicaméno sapro, sicaméno mavro sykaminia, sykamouron mavron sykaminos Morus sp Morus alba, M. nigra Morus sp. Moraceae Moraceae sico agrio^ syko sykon (fig) Ficus sp. Ficus carica Ficus carica sykéa (fig tree) Moraceae Moraceae skordo skordo skordon garlic {Allium sativum) Allium sativum L. Allium sativum L. Liliaceae Liliaceae

71 Table 4 continued Vernacular names Graecanic area Vernacular names in Greece^ Etymology of cognates Ancient Greek botany - possible (Italy) corresponding taxon, Greece according to Rohlfs (1964)^ identification^ corresponding taxon, plant family plant family sculimbri askolimpros, askolimpri skolimos edible thistle Scolymus hispanicus Scolymus hispanicus Asteraceae Asteraceae zuccho tsochos, zochos, tsionchos sonchos Sonchusl Picris echioidesl Urospermum Sonchus asper, S. oleraceus and Sonchus asper, S. oleraceus and picroides? Urospermum picroides Urospermum picroides Asteraceae Asteraceae

* Yannitsaro 1977; Tzanoudakis 1982; Lambraki 2000; Sawides 2000; Kypriotakis (personal communication); Lardos 2006 ^ Rohlfs G. (1964) Lexicon Graecanicum Italiae Inferioris; etymologisches Woerterbuch der unteritalienischen Graezitaet. Tubingen ^ Fraas 1845; Lenz 1859 [reprint 1966]; Langkavel 1866 [reprint 1964]; Hort 1916; Bois 1927; Gunther 1934; Langkavel 1866 [reprint 1964]; Lenz 1966 (Facsimile edition 1859); Brothwell and Brothwell 1969; Fragkaki 1969; Steam 1976; Baumann 1993; Raven 2000; Hondelmann 2002; Powell 2003 dypioç: in Modem Greek agrios in composition not only means wild, but is also used to describe a wild plant that bears any kind of resemblance to some plant of cultivation

72 Chapter 2 Ethnobotany 73

For example, Reichardia picroides, a well-known food plant in the Graecanic area, is

called gaddhazzida in Galliciano, Roccaforte del Greco and Roghudi, galazzida in Bova

and galatsida in Greece (Heldreich 1862; Lambraki 2000). Tozer (1889) indicated that

in Grecanico “d” has replaced “1” due to the influence of the Calabrian and other

neighbouring dialects of Italian. The fourth column in Table 4 indicates the etymology

of the vernacular names (Rohlfs 1964). Gaddhazzida has its origin in the Ancient Greek

name galactis (Dawkins 1936; Rohlfs 1964), a 6* century AD word for “milk plant”

{gala in Greek means milk). This name derived from the milky juice of the species, a

common characteristic of many Cichorioidaea (Asteraceae) and Euphorbia species. In

the Graecanic area Reichardia picroides is used for several purposes. Its young leaves

are either eaten raw as snack, or cooked as chàrta tiganimena (mixtures of wild edible

greens, cooked and fried in olive oil) (Nebel et al. In prep.). Furthermore, crushed fresh

leaves are used against toothache.

The same sound shift from “1” to “d” was also reported for maruddhaci, the vernacular

name of Hypochoeris achyrophorus and Taraxacum sp. in Galliciano. Etymologically

maruddhaci has its origin in marulion, a 6^ century AD word for a kind of chicory or

lettuce (Rohlfs 1964, p. 29). In Greece, vernacular names derived from the word

marulion are agrio marulo, which is the vernacular name for Lactuca serriola,

agriamarulia in Cyprus for Lactuca serriola and L. saligna (Zanettou-Panteli 2000), and prikomarulo, which is the phytonym of Cichorium species (Kypriotakis personal

communication) or a Crepis species in Crete (Heldreich 1862, p. 78). The morphology

and taste of different Asteraceae is very similar and vegetative plant parts, such as young

leaves of different species can hardly be distinguished. It is therefore not a surprise, that Chapter 2 Ethnobotany 74 the same vernacular name is used for different Asteraceae taxa. The same might apply to pricaddhida {Lactuca viminea) in Southern Italy, prikalida {Cichorium spp.) in Crete, and pikrallida {Taraxacum spp.) in Cyprus (Zanettou-Panteli 2000).

Another example of a cognate is masaro for wild fennel {Foeniculum vulgare ssp. piperitum) in Galliciano and agriomaratho (aypiopdpaOo) or marathos (pdpaOoç) in

Greece (Fragkaki 1969; Tzanoudakis 1982; Lambraki 2000; Sawides 2000). The word marathon (pdpaOov) was found in a list of condiments in Ancient Greek literature and is the Ancient Greek name for wild fennel (Chantraine et al. 1968). Presumably the town

Marathon (a town in Greece) was named after to the abundance of fennel in the area

(Wikipedia 2005; Marathon). Also, in the late 19* century Foeniculum officinale (under the synonym Foeniculum vulgare) was called màrathron in Greece (Heldreich 1862).

The use of fennel shoots, fennel water and fennel seed were also mentioned in an ancient record of Spanish agriculture of 961 AD (Sturtevant & Hedrick 1981).

Portulaca oleracea (purslane in English, andracla in Grecanico and andrakla in Modem

Greek) seems to have been a food plant since antiquity. Purslane was known to the

Ancient Greeks as andrachni (av0pd%vr|) as Hippocrates, Theophrastus and Dioscorides mention it (Hort 1916, p. 61 Vll.1.2 ; Bois 1927, p. 66; Gunther 1934, No 150). Today in

Galliciano, the stems and fleshy leaves are eaten fresh in salads. Also in Greece, purslane is eaten as salad, as well as cooked or baked in pies (Brussell 2004).

Additionally, physicians of Antiquity recognised the “cooling” properties of its juicy leaves (Powell 2003, p. 103). Dioscorides used purslane against headaches, stomach Chapter 2 Ethnobotany 75 discomfort, respiratory complaints, and also for the care of the eyes (Baumann 1993;

Megaloudi 2005).

In sum, phytonyms of several wild plants species with specific food use have been preserved for centuries. This might be taken as indication that these plants have been continuously used as vegetables, condiment or salads since the time Magna Graecia flourished in Southern Italy or at least since Byzantine times.

2.4.4 Food as medicines

WFPs are regarded as healthy by most of the informants, and some are also specifically used as medicine (see Table 5). Health beneficial effects were described as depurative

(help to remove toxins from the blood or internal organs), antihypertensive, digestive, diuretic, anti-arthritic, anti-diabetic, anti-cancer or to make one “feel better”. As mentioned before, bitter edible greens are perceived as being particularly healthy.

Table 5: Food plants used as medicine in Galliciano, Calabria, Southern Italy

Scientific plant Vernacular Part(s) Medicinal Family AD name name used use Asparagus asparagi Liliaceae young i diuretic acutifolius shoots Foeniculum spiro masaro Apiaceae seeds i against vulgare ssp. digestive piperitum disorders Hypochoeris costardeddhe Asteraceae leaves i blood- radicata cleansing Chapter 2 Ethnobotany 76

Table 5 continued Scientific plant Vernacular Part(s) Medicinal Family AD name name used use Lactuca viminea pricaddhida Asteraceae basal i perceived as leaves healthy because of bitterness Laurus nobilis lauro Lauraceae leaves i against stomach-ache, digestive Mentha sp. menta Lamiaceae leaves i against colds

Opuntia ficus- sico tu trucu Cactaceae flower i diuretic indica Reichardia gaddhazzida Asteraceae fresh e tooth ache picroides leaves Reseda alba gattinaria Resedaceae aerial part i perceived as healthy because of bitterness Rosa canina rosa Rosaceae young e against leaves herpes, especially for young children Silene vulgaris cavuraci Caryo- leaves i diuretic phyllaceae AD - Administration: e = external application; i = internai application Chapter 2 Ethnobotany 77

Also Rivera et al. (2005) reported that more than 50 % of the wild gathered food species

from Alcaraz, Segura and Serrania de Cuenca in Southern Spain are used medicinally and often the same plant parts are used in both preparations. In Crete, artichoke leaves and stalks are not only esteemed for their delicious taste but are also said to lower blood

sugar levels and are considered as a therapy to control diabetes type II (Leonti et a l

2006).

2.4.5 Dissemination of ethnobotanical information

An important direct outcome of this project was the public dissemination of local food knowledge. The project was not just one of academic scholarship or the industrial development of novel products but aimed at transmitting this knowledge to future generations.

A booklet with the title “Ta chorta: piante commestibili tradizionali di Galliciano” (in

English “ta chorta: traditional wild food plants of Galliciano”) written by Sabine Nebel

and translated by Peter Giovannini, was published by the School of Pharmacy University of London in 2005 (see Figure 16) and was distributed among the members of the

Graecanic communities in Calabria, Southern Italy. The booklet aims at conserving and

valuing local knowledge about wild greens, and making information about traditional

food plants and their beneficial health effects available to the local population. Chapter 2 Ethnobotany 78

Figure 16: Booklet “ta chorta: Piante

commestibili tradizionali di Galliciano” by

Nebel, Sabine (2005) School of Pharmacy,

University of London, London UK, p. 61,

Italian with English summary, foreword by

Michael Heinrich, translated by Peter

Giovannini, general index, bibliography, 2

maps, 41 coloured photographs, 21 black

and white illustrations, index of wild food

plants in Grecanico and of scientific names,

6 recipes, ISBN 0-902936-12-3.

The booklet “ta chorta” documents the use of wild edible greens by people of the ancient

Greek village of Galliciano, Calabria, Southern Italy. It is richly illustrated and provides information on the ethnobotanical field study conducted in the framework of the EU funded research project “Local Mediterranean Food Plants: Potential New

Nutraceuticals and Current Role in the Mediterranean Diet.” Furthermore, the booklet includes short monographs of the most important wild plant species including: vernacular-, scientific-. Ancient and Modem Greek name of the plant, plant family, description of the plant, habitat, part(s) used, food use and if available medicinal use/properties (see Figure 17) and some traditional recipes (see Figure 18). Chapter 2 Ethnobotany 79

Gattinaria

Grecanico; Gattinaria Greco Rohlfs: Gattinaria Kaliano: Reseda bianca Greco Antico: runivopici ‘ Greco Moderno: Oxpii pt^cvia Nome scientlfico: Resedo alba L.

Fami|lia: Resedaceae

Descrlzione delta planta; planta erbacea annuale robusta, alia fine aim. fus to eretto poco ramificalo, foglic tutte pcnnatosette. con 5 15 segmenti per lato lineari lanceolati. foglie basali addensaie in una rosetta; fieri con corolla di 5 peiali bianchi.

Habitat; muri. gbiaie, ruderi, incdti arid! e sabbiosi Gattinaria iRe»«Ja atba) e tunica crba della famiglia (0 1000m) dcUc Resedaceae chc viene consumau. Parte della planta usata; parti aeree piu lenere delta pianta

Uso allmentare. si manoia cruda con olio di oliva e pane; chorta vramcna; chorta iiganimena

ni acfii Roccaforte del Greco; Gattineria. un erba man gereccia" iRohlfs, 1964)

Uso médicinale; amara percepita come salutare

* etimologla poco chuwa »o(taoio travcriztooc Pella pwota greca (Rohlfs)

Figure 17: Example of plant monograph from the booklet ''ta chorta'' Nebel (2005)

Chorta vramena Chorta tiganimena

Erbe bollite Erbe fritte

In una peniola con acqua salata e bollenie immergere le parti In una peniola con acqua satata immergere le parti tenere tenere delle varie verdure' gia sciacquate diverse volte e far delle varie verdure' già sciacquate diverse volte e far cuocere cuocere per 20 min. circa. Sgocciolare e con dine la verduia con per 20 min. circa. olio crudo, peoetoncino piccante a piacere e poi inzuppare biscotli o pane. Sgocciolare la verdura cotta e sistemarla nella padella dove si è prima fa no imbiondire laglio in olio bollente. Fare insaporire per Qualche minute, aggiungere peperoncino piccante a ' Gaddhazzida. Gattinaria. Pricaddhida. Paparna. piacere e servire. varuddhaci. tuccho. Costardeddhe. Russeddha ed altrc ' Gaddhazzida. Gattinaria. Pricaddhida. Paparina. Erbe miste = Chorta mimmcna Maruddhaci. tuccho. Costardeddhe. Russeddha ed altrc

Figure 18: Example of two recipes: “Chorta vramena” and Chorta tiganimena’ Chapter 2 Ethnobotany 80

Unfortunately it was not possible to include the printed version in the PhD thesis, but please find enclosed the electronic version of “ta chorta” (as pdf file) on the compact disc enclosed (see Appendix 8.4). If you open the pdf file, simply press the “page down” key on the keyboard to leaf through the pages. Furthermore, the table of contents, plant indexes (p. 10 and 12), and recipes (p. 54) are tagged and linked to the appropriate page

- just click on it.

2.5 Conclusion

The Graecanic villages and specifically Galliciano are an example of rapidly changing communities where local traditions compete with modem ways of life. The present study demonstrates how the traditional consumption of wild food plant species is strongly embedded in the local culture, providing a strong link between local people and their management of the natural environment. The habit of collecting and cooking edible non­ cultivated plants is still alive among the older generation. However, it seems only a question of time before this local knowledge is lost forever. Already today, a lot of the traditional knowledge regarding food use is no longer actively used by the younger generation and is subject to many outside influences and changes. It is known, that dietary diversity undoubtedly contributes to a balanced diet in local mral communities.

Today, at the time when biodiversity is increasingly seen and valued in the context of local culture as “bio-cultural diversity” (Maffi 2001), local food knowledge represents a potential for conservation purposes. This study highlights some conceptual and methodological issues, and also practical implications. The link between biodiversity conservation and nutrition and human health, was also the subject of a recent cross­ Chapter 2 Ethnobotany 81 cutting initiative by the Convention on Biological Diversity (CBD 2005), in which the contribution of plant and animal genetic resource to dietary diversity, health and nutrition was discussed.

This comparative analysis sheds light on the historical and cultural significance of wild food plant use and its continuity in the Mediterranean and the Graecanic communities in particular. Furthermore, it contributes to an understanding of the historical development of plant use in a discipline largely relying on diachronic methods. The historical- linguistic analysis indicates, that most plant names of the culturally important wild food plant species of the Graecanic area have cognates with the same plant species used in

Greece. Etymologically these cognates have their origins in the Ancient Greek language and accordingly in Greek culture.

The cultural importance of these plant species is shown in the preservation and transmission of plant names and their associated use in the Graecanic area and Greece.

These plant species have the same culinary use in both the traditional diet of Southern

Italy and Greece and therefore, make the continuous use of a least some wild food plant

species and with it the transmission of knowledge on these plants since the time of

Ancient Greece highly likely. Consequently, this study provides indirect evidence for the history of plant use in Calabria, i.e. a region for which hardly any published

archeobotanical data could be found in a literature search in standard databases (BIOSIS previews, CAB abstracts and others) (Ferrarini & Padula 1969). It also provides a new method to better understand the development of plant knowledge in cultures relying on

the oral transmission of knowledge. The further potential of using a linguistic-historical Chapter 2 Ethnobotany 82 approach has to be explored and similar comparative analyses for other regions will hopefully further elaborate the usefulness of this method.

To re-valorise local food traditions today, a small handbook on the use of WFPs in the

Graecanic area was published (Nebel, 2005). To strengthen this local knowledge, novel curricula in schools and universities are required, as well as substantial changes in the agenda of many national food and local policy-makers in the Mediterranean (cf. Fajardo,

Verde, Rivera, & Obon, 2005). Furthermore, these data, strengthening our understanding of the cultural importance of wild food plant species and the ecozones they represent, will have a direct impact on the management of this diversity. Since such species also offer small-scale business opportunities locally, they are more likely to be seen as a valuable asset to be preserved (Pieroni et a l 2002b; Verde et a l 2003; Pardo de

Santayana et a l 2005a; Pieroni et al 2005; Tardio et al 2005). Chapter 3 Socionutritional study 83

3 Socio-nutritional study in six rural areas of the Mediterranean

Socio-nutritional study on the consumption of traditional food plants in a Graecanic community in Calabria, Southern Italy and five other rural areas of the Mediterranean.

Signora Ciccia, Galliciano 2002 Chapter 3 Socio-nutritional study 84

3.1 Introduction

Food use is changing very fast on all continents. In industrialised countries, there is a

considerable rise in expenditure for convenience food (pre-prepared or ready made dishes). At the same time, nutraceuticals and functional foods are a rapidly growing

segment of the market (Hardy 2000). Concurrently, there is a dramatic and un-revocable

loss of “local knowledge” regarding food use, which forms the basis of many cultural traditions (traditional food knowledge). These and other changes, such as reduced physical activity or increased longevity, result in novel health risks for the populations in

European countries and beyond (Popkin 2004).

Traditional food knowledge (TEK) is strongly influenced and determined by socio­ economic and cultural parameters, as well as religion and history (Johns et a l 1994;

Shatenstein & Ghadirian 1998). All food is part of human’s everyday experience and the way it is perceived and classified forms the basis for food use in a culture. Around the

Mediterranean, a multitude of cultures, religious beliefs, écologie backgrounds and historic developments resulted in many diets which share a lot of elements but also

revolve around distinct local or regional traditions (Nestle 1995). “Local food” as part of

local traditions, is based on ingredients, which are gathered, grown or produced locally

and prepared into dishes, which often represent local specialities (Heinrich et a l 2005b).

Vegetables and salads comprising wild greens are especially important as local foods.

WFPs have been an element of Mediterranean dietary traditions for millennia. The

tradition of consuming WFPs still plays an important role in the traditional livelihood of Chapter 3 Socio-nutritional study 85 rural people in the Mediterranean, despite the primary reliance of agricultural societies on crop plants. In the past decade, a few studies have systematically analysed the consumption of WFPs in the Mediterranean area from an ethnobotanical perspective

(Forbes 1976' Paoletti èt al: 1995; Bônet & Valle's 2002; Guafrera 2003; Pafdd de

Santayana et a l 2005b; Pieroni et al. 2005; Rivera et a l 2005; Tardio et al 2005; Leonti et al 2006; Tardio et al 2006). AU these studies clearly demonstrate that these local foods still represent a relevant part of traditional Mediterranean diets. In some cases,

WFPs are basically considered as famine food or as the food of the poor, but in many

European countries the acculturation and globalisation have been accompanied by a revaluation of local identities and traditions, including traditional cuisines. Additionally, the nutritional potential of non-cultivated vegetables native to the Mediterranean and the

Near East, and their potential health benefits have been recognised as an important area of research (Guil Guerrero et al 1998; Trichopoulou et a l 2000; Pieroni et a l 2002a;

Couladis et a l 2003; Tarwadi & Agte 2003; Zeghichi et a l 2003).

However, assessing changes in local diets are difficult since in many instances

“traditional” or local dietary patterns have already disappeared and often no quantitative data on food consumption in traditional societies are available (Helsing 1995; Tumino et a l 2002). Consequently, systematic investigations regarding traditional food culture are urgently required. This applies especially to those areas which, for geographical and historical reasons, remain relatively isolated and where traditional food use practices are still alive, but risk to disappear in the future due to industrial or other development. Chapter 3 Socio-nutritional study 86

In the interdisciplinary study in hand, ethnobotanical and nutritional anthropological methods were used to assess the diversity and the role of WFPs in local nutrition and cuisine. The study identified plants traditionally consumed in rural communities in

Southern Italy (see Chapter 2), quantified their consumption also in relation to the everyday diet, assessed the local perception of health benefits or risks associated with wild food plant consumption and, equally important, added new value to local food products which have been used for many generations.

3.2 Aims and Objectives

This socio-nutritional study focused on a detailed understanding of the social, economic, and educational characteristics, anthropological factors, health status (quality of life) and overall dietary patterns in six rural areas of the Mediterranean, in order to:

• describe the social, economic, and educational characteristics of each area and

assess their association with decisions regarding the consumption of foodstuffs

• identify psychological and anthropological factors, which are important to

consumers in their choice of specific plant dietary components

• examine local beliefs regarding the health benefits linked with the consumption

of wild gathered food plants

• study the consumption of WFPs in the local dietary pattern

• identify dietary patterns that might be important in the prevention of the ageing

related illnesses (ARIs) Chapter 3 Socio-nutritional study 87

Research Questions

What is the role of wild gathered food plants in local diets around the Mediterranean?

How often are wild gathered food plants consumed in rural regions of the

Mediterranean? What is the local perception of the consumption of wild gathered food plants regarding health benefits and risks? What factors influence the consumers in their

choice of specific foods? Is the local diet adherent to the healthy “Reference

Mediterranean diet”?

3.3 Methodology

In the following sections, methodological aspects of the socio-nutritional study of the

LFN project are presented. The authors contribution to this socio-nutritional study was: designing the socio-nutritional questionnaire, conducting the interviews in Galliciano in

Southern Italy in 2003, developing the SPSS data file, inputting the gathered data from both South Italian villages (Galliciano and Castelmezzano) into SPSS and finally

conducting the overall analysis of the socio-nutritional study (including all study sites;

with emphasis on Southern Italy). All data relevant for Galliciano are highlighted in

grey.

The following sections include details on the overall framework of the socio-nutritional

study, the selection of the study areas, the questionnaire used in the socio-nutritional

study, the pilot study, the sampling and recruitment of households, as well as

information on the data processing and analysis. Chapter 3 Socio-nutritional study

3.3.1 Sampling and recruitment

3.3.1.1 Sampling strategy

The socio-nutritional study was conducted in three regions of the Mediterranean: in

Southern Italy, South-Eastern Spain (SE Spain) and Crete (Greece) by researchers of the

LFN consortium. In each of these countries, two areas with either high or low consumption of wild gathered food plants were selected, guided by data gathered during previous ethnobotanical studies in the three countries, in order to compare the characteristics of consumers and non-consumers (or low consumers) of locally gathered foods.

These sub-samples in SE Spain and Greece were selected within the same study areas to ensure as far as possible that they had broadly similar characteristics and lifestyle factors. In the Italy, the socio-nutritional study was conducted in the same villages, in which the ethnobotanical studies were conducted in 2002. A main criterion for choosing these two areas was their different socio-cultural pattern. Castelmezzano is located in the inland part of the Region Lucania and the culture is strongly influenced by a typical mountain culture. Inhabitants are ethnically South-Italians, who have relevant traces of

Norman, Swabian, and Spanish domination (10* -16* centuries) (Pieroni et a l 2005).

Food habits are still depending on swine husbandry (high consumption of pig fat).

Nowadays, most of the youngest inhabitants work in factories in Potenza (regional capital). Galliciano, on the other hand, is located in the South of the Aspromonte mountain areas, very close to the Ionic coast, in Southern Calabria, one of the regions with the lowest income per person in Italy. The inhabitants of Galliciano are, as Chapter 3 Socio-nutritional study 89 compared to the surrounding Italian population, characterised by their own Greek dialeet

Grecanico, culture and history as an ethnic and linguistic minority (ethnic Greek; see

Chapter 2.1.1 Ethnographic background of Graecanic area). Still today, agro-pastoral activities, such as olive tree cultivation, are part of the traditional livelihood in the area.

In total, the seleeted study areas eomprised five regions (Castelmezzano and Gallieiano in Southern Italy; Salobre and Viveros in South-Eastern Spain; Crete A in Greece) with population sizes less then 1000, and one larger area with 5000 inhabitants in Greece

(Crete B). The characteristics of the study areas are given in Table 6.

Table 6: Overview table of number of inhabitants and households in the selected study sites of the Mediterranean

Number of Numbers of Consumption of inhabitants households WFPs Southern Italy Gallicianô (Calabria) Approx. 80* 16* high Castelmezzano (Lucania) 838** 352** high South -Eastern Spain Salobre 622*** 98*** high Viveros 564*** 138*** low Greece Crete A 677 85 high (Avdou, Amariano) (414 + 263)

Crete B 4991 (1940 + 140 low

(Anogia, Zoniana, Livadia) 1578 + 1473) * data source: project field worker 2002 ** data source: munieipal statistical office 2002 ***data (relating to 1998) from National Institute of Statistic, 2002 Chapter 3 Socio-nutritional study 90

To fulfil the objectives of the study, a representative sample from the study areas was

required. This enabled the investigation of characteristics and health perspectives of the population of the selected areas. Households, the unit of interest, were randomly

selected, to obtain representative samples in each area. All members of the household

(older than 18 years), who were available, were interviewed with the same interview

schedule.

3.3.1.2 Sampling procedure

In the Castelmezzano, SE Spain and Crete A and B the households were a systematic

sample in that every n‘^ household was selected and invited to participate the basis of a

street map. In the case of the Graecanic area, in which the author S Nebel conducted the

socio-nutritional survey, the village of Galliciano includes 16 permanent households

each with between one and eight inhabitants. From the households which gave consent to this study, eight households were randomly selected using the Research Randomiser

on the internet (http://www.randomizer.org/form.htm). Oral consent was obtained prior

to the commencement of each interview with household members.

To obtain a representative sample it was important to maximise the response rates. In

cases in which no response was obtained, households were re-visited at different times

of the day. For cases in which the members of the household choose not to participate, a

further household was selected according to the sampling procedure for that area. Chapter 3 Socio-nutritional study 91

3.3.2 Data collection

3.3.2.1 Development of the socio-nutritional questionnaire

A socio-nutritional questionnaire on the consumption of traditional food plants and its relation with social, economic and anthropological factors has been developed by the author together with Prof. Michael Heinrich, Prof. Felicity Smith (School of Pharmacy) and Dr. Andrea Pieroni (until 2002 as post-doc at the School of Pharmacy in London, now University of Bradford). The questionnaire was developed for household members aged over 18. It was structured in four sections, including open and closed questions (see

Table 7; see Appendix 8.2 for complete questionnaire in Italian)

Table 7: The structure of the socio-nutritional questionnaire used in the “Local food - Nutraceuticals” project

Section 1 Information on WFPs that are Which products are consumed consumed in the household Frequency of consumption Perceptions regarding the nutritional and health benefits Perceptions regarding any risks to nutrition and health Factor which influence the choice of food products Section 2 Aspects of lifestyle and diet Smoking relevant to health Physical exercise Diet: data collected using a Food Frequency Questionnaire FFQ (established tool for nutritional assessment) Chapter 3 Socio-nutritional study 92

Table 7 continued

Section 3 Information on health status and • EuroQol (EQ-5D): established personal characteristics of tool for the measurement of respondent health status • Data on sex, age, education, occupation and residency Section 4 Other people in the household • Data related to household structure (number of people living in a household; age and sex of household members) for the identification of individuals to be interviewed

Data were collected in structured interviews with adult members of households who were available at the time. Household members were defined as those residing at the time of the study and normally cook and /or eat together one meal an average of one meal daily. Younger members of the community (18 to 30 years old) were included in order to evaluate whether local knowledge about food plants was passed on in the community (intergenerational factor).

In section one, the intake of wild gathered plants was assessed by asking the question:

“how often do you eat [name of plant] when it is in season?” Response options ranged from “never” to “most days”. Any health beneficial effects or health risks associated with wild gathered vegetables were assessed with further questions: “Does this [names of the plant] have any helpful/beneficial effect on your health? If yes, please describe these effects in detail”. “Are there any health risks or possible adverse effects associated Chapter 3 Socio-nutritional study 93 with this food? If yes, what are these?” Prompts were used to gather more information on possible health risks, such as: “Do you think it affects your general health/life expectancy? Do you think it has an effect on any particular condition or symptoms, and if so which?”

The question regarding food choice comprised eight factors that may influence food choice, which had to be rated as “not important”, “sometimes important” or “very important”. It is known that food choice is influenced by many interrelating factors ranging from biological mechanisms and genetics on the one hand, to social and cultural factors on the other (Shepherd 2002). After the interviewee had rated the different prompted factors, they were asked to quote the two most important and the two least important factors influencing their choice of food.

Aspects of lifestyle relevant to health were assessed with three questions regarding smoking, physical activity/work and other exercises for leisure. General aspects of diet were covered with an extensive food frequency questionnaire (FFQ, see Appendix 8.2), asking how many portions of different food items they have eaten in the last twelve month. The FFQ was recoded afterwards to evaluate “the number of portions of specific food items per week”. The FFQ was initially developed by the Greek partners

(Department of Nutrition and Dietetics, Harokopio University in Athens) of the LFN project, translated into Italian and Spanish, validated and tested upon small samples to make sure that it was comprehensible. FFQ is a widely used tool to investigate customary food intake over extended periods of time (Marks et al. 2006). A tendency for respondents to food frequency questionnaires to under-report consumption of foods Chapter 3 Socio-nutritional study 94 perceived as unhealthy (e.g. butter, processed meat) and to over-report consumption of foods perceived as healthy (e.g. fruit and vegetables) has been reported (Salvini et al

1989). These biases affect the usefulness of food frequency questionnaires in the assessment of total dietary intake for individuals. However, they would only be important for the analyses reported if different individuals show different biases.

Information on health status was obtained with the EuroQol (EQ - 5D) which was included in section three of the questionnaire. EQ-5D is an established measurement of health status and quality of life developed by the EuroQol Group in 1987 (The EuroQol

Group 1990). This tool was selected, because it was developed and validated on an

European level, and was available in Italian, Spanish and Greek. It is a simple, generic health related quality of life (HRQoL) questionnaire with two principal measurement components. The first descriptive system defines health in terms of five dimensions

(5D): 1. mobility, 2. self care, 3. usual activities (work, study, housework, family, or leisure), 4. pain or discomfort and 5. anxiety or depression. Each dimension is subdivided into three categories, which indicate whether the respondent has no problem

(= 1), a moderate problem (= 2), or an extreme problem (= 3). The respondents select one level in each dimension, thereby defining their health state in terms of a five figure code (Badia et a l 1998). For example, the state 11111 represents no problems in any of the above mentioned dimension, whereas 33333 stands for severe problems in all dimensions. Combinations of these dimensions define a total of 243 health states. The second part of the EQ-5D consists of a vertical, 0-100 visual analogue scale (VAS), where 0 denoting the worst and 100 the best imaginable health state to record the interviewees perception of their overall health. The EQ-5D therefore provides Chapter 3 Socio-nutritional study 95 quantitative information about the respondent’s health state. Two more questions regarding health problems were asked: firstly participants were asked to rate their health during the past four weeks ranging from excellent to poor, and the secondly to state any health problems diagnosed or treated by a doctor in the previous year.

Furthermore, descriptive data related to the population of each of the study site were gathered by the researcher. This descriptive data included: population of each community/ village, number of households, household size, population age structure, ethnic groups in each area (e.g. ethnic origin, language spoken), employment patterns, any other social, economic, cultural indicators or descriptors.

To summarise, the following information was obtained in structured interviews with household members:

Information on used tool in the questionnaire wild food plant consumption open and closed questions dietary assessment food frequency questionnaire (FFQ) health status EQ-5D personal information of household closed questions members

The wording of questions was adhered by the interviewer throughout the interview and prompts were used to obtain additional information regarding open questions. Chapter 3 Socio-nutritional study 96

3.3.2.2 Approval from local research committees

In each of the six study sites, the national/local regulations were followed and approval was obtained prior to the commencement of the data collection in each site.

3.3.2.3 Study procedures

Questionnaires, information leaflets and consent forms were translated into the principal

(national) languages of each of the study sites (Italian, Spanish, Greek). To ensure reliability and validity of the translation, all documents were translated back into English by a second person. Any discrepancies were addressed by the research team and amendments checked again by independent back-translation.

To ensure the feasibility of the study procedures and that the interview schedules result in the collection of the data required to meet the study objectives, a pilot study with a few interviews was arranged and conducted in each country. For example, the length and procedures of the interview had to be acceptable and all materials had to be comprehensible. All questions in the interview schedule were assessed for difficulty of interpretation, missing data, clarity of responses, unclear responses.

3.3.3 Data processing and analysis

Coding frames were devised for the questionnaire data. The data were coded and entered into SPSS to enable statistical analysis. First of all, the data from all study areas were analysed. This included the generation of frequency data and summary statistics.

Statistical comparisons were undertaken using principally non-parametric procedures: chi-square tests, Mann-Whitney U, Kruskal-Wallis, Spearman’s rank correlation. In all Chapter 3 Socio-nutritional study 97 cases a p<0.05 was taken as conferring statistical significance. The SPSS data file was constructed centrally and the data from each of the study sites entered. On receiving the data from each interviewer, the consistency of the coding for all data sets was checked.

Secondly, the data from Galliciano, Southern Italy were analysed in more depth, to gain a better understanding of the Graecanic area, which was the main focus of this thesis.

However, due to the small size of the Graecanic community Galliciano (16 households) only 22 questionnaires in seven households could be administered, allowing only limited statistical analysis including the generation of frequency data and summary statistics with SPSS version 13.0.

3.4 Results and Discussion

Data collection was done in April 2003 and completed in May 2003. The socio- nutritional study was carried out simultaneously by researchers of the LFN Consortium in six different Mediterranean regions.

3.4.1 Recruitment settings: households, response rate, and interviewers

In total, 321 interviews were conducted in the six study areas in 184 households (for details of all study sites see Table 8). In Galliciano, seven households were randomly selected. This resulted in a sample of 12 women (55 %) and 10 men (45 %) with an average age of 46 years (from 18 to 97 years old). There was a good response rate of

85 %, with a total of 22 completed interviews. Chapter 3 Socio-nutritional study 98

Table 8: Sample size (number of households included in the study), total of interviews conducted, response rate in all six study areas and the interviewers

Number of households Total Response Interviewers inch in the interviews rate study Southern Italy Galliciano 7 22 85% S. Nebel

Castelmezzano 3 7 3 9 77% A. Pieroni SE Spain

Salobre 2 9 51 93% C. Inocenico / A. Verde

Viveros 35 71 88% C. Inocenico / A. Verde Crete (Greece)

Crete A 41 6 4 81 % G. Felliou / S. Sidossis

Crete B 35 7 4 83% G. Felliou / S. Sidossis Total 184 321 84%

A response rate was calculated regarding the proportion of household members who agreed to participate in the socio-nutritional study. There was a good response rate in all study areas between 77 % and 93 % (see Table 8). The reasons for not participating in the study were lack of time (especially male members of the household) and physical impairment. On average it took about 29 minutes to interview one member of a household (average of 29 minutes; ranges = 10 to 75 minutes; N = 321).

3.4.2 Characteristics of study sites and respondents

Three hundred and twenty-one household members were interviewed; of whom 194

(61.4 %) were female and 122 (38.6 %) were male (5 answers are missing). The average household size varied from 1.8 to 4.43 persons per household. Chapter 3 Socio-nutritional study 99

3.4.2.1 Age structure

The oldest participant was 97 years, the youngest 18 years. The mean age of the respondents was 59.6 years. The following box plot (see Figure 19) shows the age distribution in the six study areas. A box plot provides an excellent visual summary of many important aspects of a distribution. The box stretches from the lower hinge

(defined as the 25th percentile) to the upper hinge (the 75th percentile) and therefore contains the middle half of the scores in the distribution. The median is shown as a black line in the box.

What is your age In years?

1 0 0 -

Italy- Spain- Spain- Greece - Greece- Galliciano SalobreCastelmezz Viveros Crete A Crete B

Figure 19: Box plot of the age distribution in the six study areas (Southern Italy, South-Eastern Spain and Crete)

The age structure in Italy, Castelmezzano; Spain Salobre; Spain Viveros and Crete B is very similar, while in Galliciano and Crete A the age structure is slightly different. In Chapter 3 Socio-nutritional study 100

Galliciano the percentage of younger people is higher than in any other study area, whereas in Crete A the average age is significantly older. Furthermore, the age structure is significantly (p < 0.01) different between the high and low consumer areas in Greece.

3.4.2.2 Ethnicity

In all study regions, except for Galliciano, one ethnicity was recognised per region: in

Castelmezzano they described themselves as Southern Italian, in Salobre and Viveros as

South Eastern Spanish and in Crete A and B as Greek. In Galliciano, Southern Italy,

63 % described themselves as “Greek Italian” and 37 % as “Southern Italian”. In general, it was observed that the older generation (age older than 55) described themselves as “Greek Italian”, whereas the younger generation (age younger than 25) named themselves “Southern Italian”. As the village is in a state of transition, the result shown above does not come as a surprise. Younger people have to migrate from the traditional hamlet in the mountains to the villages along the coast or abroad, as there is no longer a school or work to make a living in the village. Some recent initiatives in the village in “reviving traditions” gives hope, that the younger generation remember their origin, their traditions and their ancestral language Grecanico, which is nowadays only spoken by the older inhabitants. Therefore the younger inhabitants of Galliciano are aware of their ethnic origins and if in the questionnaire an option would have been given to answer with “Southern Italian with Greek origin” (Italiano con origine Grecanico) the results would look different.

3.4.2.3 Education and employment status

The average years of full-time education varied from 4.6 to 7.0 years in the six study areas. The proportion of people who did not go to school is the highest in Spain (37 and Chapter 3 Socio-nutritional study 101

29 %), followed by Greece (7 and 5 %) and Italy (5 %). Most of the household members described their employment status as being housewife, retired or working full-time. The proportion of participants seeking work or being unemployed was significantly higher in

Galliciano (5 out of 22) than in the other study sites. This might be explained by the fact that migration is very high in Galliciano (Region of Calabria) and that younger inhabitants of the village often seek seasonal work abroad (see Table 9).

Table 9: Current employment status of household members in the six study areas (numbers in %)

In edu­ working working house­ un­ N retired cation full-time part-time wife employed

T otal 320 2.5 % 20.3 % 6.3 % 43.1 % 2.8 % 25.0 % Southern Italy Galliciano 22 9% 4% 5% 36% 23% 23% Castelmezzano 39 - 25.6% 10.3% 28.2% - 35.9% SE Spain Salobre 51 4% 29% 4% 41 % - 22% Viveros 71 6% 17% 4% 56% - 17% Crete (Greece) Crete A 64 - 2% 3% 48% 3% 44% Crete B 73 - 36% 11 % 37% 3% 13%

3.4.2.4 Sex, age of respondents

In all study areas more women participated in the interview than men (see Table 10).

Overall, slightly more than half of the interviewees (54 %) had the responsibility in deciding what foods were eaten in the household, the other half (45 %) did not have any responsibility and only 8 % of the interviewees did share the responsibility for deciding what foods are eaten in the household. In Galliciano the responsibilities were more often Chapter 3 Socio-nutritional study 102 shared with other members of the household (32 %) than in the other study sites. In several households in Galliciano, mothers shared the responsibility for household matters with their daughters, as they had to take over household matters, when the parents were engaged otherwise, like working in home gardens.

Table 10: Sex, Age (mean and SD) of respondents in the six study areas and analysis of responsibilities for deciding what foods are eaten in the household.

Mean age of Responsibility for Sex of respondent deciding what foods are respondent lyearsj eaten in the household Female Male Female Male Yes Shared* No 60 59 38 Total N 61 % 39% 54% 8% (±19) (±17) % Southern Italy G allicianô 22 54% 46% 46 47 23% 32% 45% Castelmezzano 34 76% 24% 5 55 92% 3% 5% SE Spain Salobre 51 59% 41 % 62 61 55% 4% 41 % Viveros 71 62% 38% 59 61 51 % 6% 43% Crete (Greece) Crete A 64 63% 37% 73 69 56% 14% 30% Crete B 74 57% 43% 59 52 45% 4% 51 %

(e.g. mother and daughter decide together)

3.4.3 Wild food plant consumption

In general, between the regions selected in Southern Italy, Spain and Greece, there was a significant difference in wild food plant consumption (see Table 11). In all study sites wild gathered food plants were consumed regularly when in season, which is Chapter 3 Socio-nutritional study 103 approximately from December until May. They are very valuable as vegetable substitute in early spring, as they are available several weeks before the garden varieties. The broad range of species which is used in the traditional cuisine, is reflecting the seasonal aspect of gathering, as well as the species’ taste. The analysis showed that, on average, the participants in Galliciano consumed three portions (1 portion equals 1/2 cup, cooked) of WFPs a week, while participants in Castelmezzano, Viveros (SE Spain) and Crete B consume about one portion per week. In the high consumer region in SE Spain (Salobre) participants consumed about two portions a week and in Crete A about two and a half portion per week.

Table 11 : Consumption of wild food plants (results of the FFQ) linked with the frequency of wild food plant consumption (section 1 of the questionnaire)

Mean portion Frequency of wild food plant WFPs per week consumption - how often do you eat (1 portion = 1/2 WFPs when they are in season? cup, cooked) once a week at least once N to once a less often (FFQ) a week month Southern Italy Galliciano 22 3.0 (±2.4) 9% - 5%r % Castelmezzano 39 1.2 (±1.8) 15% 44% 41 % SE Spain Salobre 51 1.9 (± 1.4) 12% 67% 21 % Viveros 71 0.8 (± 0.7) 13% 55% 32% Crete (Greece) Crete A 64 2.5 (± 2.4) 8% 64% 28% Crete B 74 0.7 (± 1.7) 15% 51 % 34%

frequency of wild food plant consumption (how often WFPs are consumed during a week or month) was significantly different between Galliciano - Castelmezzano Chapter 3 Socio-nutritional study 104

(p < 0.001) and Crete A - Crete B (p = 0.004). In Galliciano the majority of the people consumed wild foods “at least once a week”, while in Castelmezzano 43 % of the interviewees consumed WFPs only “once a week to once a month”. This showed that in the “high consumer” area in Crete A and Galliciano, WFPs were consumed more often

(frequency) and in larger quantities (portions per week). With only the analysis of the

Food Frequency Questionnaire this difference would not have been visible. In Spain, there was no significant difference in the frequency of consumption between the low and the high consumer area.

Some further analysis of the data from Galliciano showed, that despite the expectation that mostly the elderly eat wild greens, there is no significant difference between the age groups (younger and older than 45 years) nor between sexes, regarding the consumption of WFPs. About a third of the interviewees (32 %) consumed WFPs “most days”, while

55 % eat them “at least once a week” and only 13 % have them “once a week to once a month or less often”. However, the older interviewees in Galliciano unanimously agreed that today far fewer wild gathered food plants were consumed than in previous decades.

Especially in times of famine, the consumption of wild gathered food plants was much higher. And still today in many areas of the Mediterranean the consumption of WFPs is often linked to scarcity and “food of the poor” (Pardo de Santayana et al. 2005b). A progressive decline in the importance of gathered foods was already reported in 1997

(Van der Meer 1997), and yet numerous studies illustrate that rural people in many parts of the world continue to include a multitude of wild edible plants in their diet, and that these make important contributions to macro- and micronutrient intake (Scoones et al.

1992; Grivetti & Ogle 2000). Chapter 3 Socio-nutritional study 105

Local diets: results o f the Food Frequency Questionnaire FFQ

The Mediterranean dietary pattern is characterised by a high intake of vegetables, legumes, fruits and nuts, cereals, a high intake of olive oil and low intake of saturated lipids, a moderately high intake of fish (depending on the proximity to the sea), a low-to- moderate intake of dairy products (and then mostly in the form of cheese or yoghurt), a low intake of meat and poultry and a regular but moderate intake of alcohol, primarily in the form of wine and generally during meals (Trichopoulou & Vasilopoulou 2000;

Trichopoulou 2001; Trichopoulou et al 2003). However, a major nutritional concern in the Mediterranean countries is the loss of the traditional Mediterranean dietary pattern

(Trichopoulou 2001; Moreno et al 2002).

In the six study areas of this study, differences in dietary habits were observed. The dietary pattern in Salobre, and Crete A were closer to the Mediterranean pattern (high fruit and vegetables consumption, and low red meat) than the other regions. The comparative analysis of the FFQ showed that A.) Greece was the country with the highest grain product and vegetable consumption; B.) Spain had the highest fruit and wild green consumption; C.) consumption of legumes, as well as meat, were alike in all the regions and D.) the fish consumption was highest in Spain.

Compared to the “typical” Mediterranean dietary pattern described originally, the data from Italy showed a poor compliance, meaning a trend to Westernisation of the dietary habits, which was also reported in other rural areas in Italy by Alberti-Fidanza et al.

(1999) and Barbagallo et al. (2002). A detailed analysis of the data from Southern Italy

(Castelmezzano and Galliciano) showed that the consumption of legumes and wild Chapter 3 Socio-nutritional study 106 greens was significantly higher (P < 0.001) in Galliciano, than in Castelmezzano. In

Italy, a progressive increase of consumption of meat, fats, milk and dairy products and sugar has been reported especially in the north, but nutritional habits also changed in southern parts of the country (Ferro-Luzzi et al. 1984; Alberti-Fidanza et al. 1994). The data from the different counties were analysed separately, since each country has its own

“Reference Mediterranean Diet” (Alberti-Fidanza et al. 1999).

Fruit and vegetables are important components of a healthy diet, and their sufficient daily consumption could help prevent major diseases, such as cardiovascular diseases and certain cancers (Eaton 2003). Still, most populations are not consuming nearly enough, according to the F AO/WHO Expert Report on Diet, Nutrition and the

Prevention of Chronic Diseases (WHO 2003). In the six study areas of this research project, the total fruit and vegetable consumption was more than the generally recommended five servings a day (five servings equal 400 g of vegetables and fruits per day). Noteworthy was the contribution of WFPs to the total fruits and vegetables consumption (see Table 12). Chapter 3 Socio-nutritional study 107

Table 12: Total consumption of fruits and vegetables (including WFPs) per day.

Consumption of x Contribution of WFPs N portions of fruits and to the total fruit and vegetables per day vegetable intake Southern Italy Gallicianô 22 9.1 28% Castelmezzano 39 6.3 3% SE Spain Salobre 51 11.5 22% Viveros 71 9.5 11 % Crete (Greece) Crete A 64 8.7 14% Crete B 74 8.5 4%

This socio-nutritional survey supports also the thesis that the term “Mediterranean diet” is a misnomer. Actually, there is not just one Mediterranean diet, but in fact many

Mediterranean diets (Simopoulos & Visioli 2000). This is not surprising since the countries along the Mediterranean basin have different economical and cultural traditions and politically the Mediterranean region is anything but homogeneous. In addition, the basic conditions for food production varied considerably around the

Mediterranean shores (Helsing 1995). The population supplemented their diet with locally grown foods since millennia, producing a diverse variety of regional varieties and specialities. However, despite the given differences, the Mediterranean diets still share several key elements and health benefits. Chapter 3 Socio-nutritional study 108

Oil used for cooking, salad dressings or sauces

In all study sites olive oil was the favourite oil for cooking, salad dressing or sauces.

Other oils used for cooking were lard in Castelmezzano (44 % of the informants) and sunflower oil in Galliciano (65 % of respondents). For the preparation of salad dressings nearly all informants used exclusively olive oil and only 3 % of the interviewees in

Viveros (SE Spain) used sunflower oil instead of olive oil. When analysing oils used to prepare meat or fish sauces, only the respondents in Greece used olive oil exclusively. In the Castelmezzano participants mentioned not only olive oil, but also lard, sunflower oil or butter for sauces. Interestingly in Viveros 35 % of the interviewees used sunflower oil instead of olive oil to prepare sauces.

In general it can be said, that in the Mediterranean basin, olive oil, along with fruits, vegetables and fish, is an important constituent of the diet, and is considered a major factor in preserving a healthy population (Galli & Visioli 2001; Perez-Jimenez 2005). A diet, rich in virgin olive oil, improves the major risk factors for cardiovascular disease, such as the lipoprotein profile, blood pressure, glucose metabolism and antithrombotic profile (Trichopoulou et a l 2003). It has been shown that in countries with a diet closely following the Mediterranean diet, and where olive oil is the principal source of fat, cancer incidences rates were lower than in northern European countries (Willett 2001).

3.4.4 Perception regarding health benefits or risks

In all six regions WFPs were regarded as healthy by 66 % of the informants participating in the socio-nutritional study (N = 321), while 33 % did not know whether wild food plant have potential health benefits or not. Beneficial effects were described as Chapter 3 Socio-nutritional study 109 depurative, antihypertensive, digestive, diuretie, anti-cancer or to make one “feel better’

(see Figure 20).

Beneficial effects were described as: Do plants have any helpful/beneficial effect on your health? Depurative (help to remove toxins from the blood and internal organs) Antihypertensive 33.0% Digestive □ Yes Diuretic ■ No Laxative □ Don't know “feel better”

65.8 % Anti-arthritic Anti-diabetic Anti-cancer “Clearer thought”

Figure 20: Perception of WFPs regarding any helpful/beneficial effect on health by local people in the six study areas in the Mediterranean

As mentioned before, hitter edible greens are perceived as being particularly healthy

(see Section 2.4.4. Food as medicine). The consumption of WFPs is highly relevant for health, as they often contain higher amounts of bioaetive compounds than plants that have been under cultivations for many generations (Lionis et al. 1998; Stepp 2004).

Trichopoulou et al. (2000) showed, that eight wild green species consumed in Crete have a very high flavonoid content when compared with cultivated fresh vegetables, fruits and beverages commonly consumed in Europe. Furthermore, wild gathered greens and fruits are more fibrous and contain higher concentrations and a greater diversity of secondary compounds. These secondary compounds are chemicals with a wide range of potentially physiological and pharmacological interactions including toxic, antimicrobial, anti- Chapters Socio-nutritional study 110 inflammatory, antioxidant, diuretic, laxative, and more properties. In this research project, the antioxidant activity of several WFPs was analysed in in vitro assays (see

Chapter 4. Antioxidant activity of local food plants), and it could be shown, that several of the investigated WFPs showed high or moderate activity, supporting the local beliefs regarding health beneficial effects.

In the perception of most informants (88 %), WFPs possessed no health risks or possible adverse effects, whereas 9 % of the informants did not know. Only 3 % of them associated some risks with wild food plant consumption. These were described as “to be avoided by people suffering gastritis or colitis”, allergy or toothache. The informants clearly mentioned that these risks were related to the dose. One informant from

Galliciano said: “if I eat a bit [of wild food plants], I enjoy it - if I eat too much, it gives me stomach-ache”. In general, there is not enough bioscientific (phytochemical or pharmacological) information available to assess potential health risks of the dishes used in the Mediterranean area. Further phytochemical research is needed to ensure the safety of these local food plants.

3.4.5 Factors influencing the choice of food products

In Italy and Spain the most important factor influencing the choice of food products was

“to maintain a healthy diet” (42 %, and 47 % respectively), followed by “taste preferences of my family or friends” (42 % [Italy] and 65 % [Spain]). Interestingly in

Greece, the most important reason why certain food products were chosen, was the own preference “Food that I like - my preference” (51 %) followed by “taste preferences of my family and friends” with 21 %. Surprisingly in all areas only once “local traditions” Chapter 3 Socio-nutritional study 111 were mentioned to influence the choice of food products spontaneously. Furthermore, the analysis showed, that, factors not important in the choice of food were “time and convenience when shopping or gathering” in Italy and Spain (70 %, and 76 % respectively) followed by the “difficulty/ease of preparation”. In Greece “storage” and

“difficulty/ease of preparing” (43 % and 23 %) were the least important factors. No significant difference (Pearson were found when analysing the data regarding the sex of the respondents, age, the level of education, employment status, or the level of wild food plant consumption.

The results show that sensory perceptions of food, like taste, are very important factors influencing food choice. As mentioned in Chapter 2, taste is a key criterion for perceiving, categorising and characterising food plants in general (Grivetti 1981; Johns

1986; Nebel 2001). The ethnobotanical study showed, that gatherers of WFPs in

Galliciano, for example, pay attention to mix bitter and sweet herbs together to assure a balanced taste of the dishes. Bitterness plays also a very important role in the local perception of health. Therefore, it does not come as a surprise, that both taste and health are some of the most important factors influencing the food choice.

3.4.6 General aspects of lifestyle and health

3.4.6.1 Smoking and physical exercise

The majority of the interviewees (81 %) were non-smokers. Of the smokers, three quarters (75 %) were male. The mean of “actual number smoked as reported by respondent” varied from 4 to 80 cigarettes per day (mean = 27 ± 18). Furthermore, the analysis showed that most of the respondents (68 %) did physical exercise like farming. Chapter 3 Socio-nutritional study 112 gardening, housework, chopping firewood, lopping olive trees and many other activities

(see Figure 21 and Figure 22). Only 49 % of the respondents did exercise for leisure, including sports, walking, hunting or picking greens.

m k

Figure 21 : Signora Maria collecting Figure 22: Girls carrying oranges from the

olives (white bucket) and wild greens homegardens back to the village

(blue bucket) in her olive grove, Galliciano

Galliciano, Southern Italy

3.4.6.2 Health status of household members (EQ-5D)

Only 14 % of the health states generated by the EQ-5D descriptive system were selected

(35 out of 243). Thirty-three percent of the overall sample rated their health state as Chapter 3 Socio-nutritional study 113

11111. Only two respondents in Greece selected the most dysfunctional health state

(33333). Low mean visual analogue scale (VAS) values corresponded to a poorer health state as reported on the EQ-5D descriptive system.

Table 13 shows the percentage of individuals reporting problems in each of the EQ-5D dimensions and the mean VAS values according to the study sites. Pain/discomfort was the most frequently declared problem (50 % of individuals) and self-care the least frequently declared (7 % of individuals). Anxiety/depression problems were reported by

40 % of the population and the mean VAS score for the overall health status was 69.0

(SD= 18.4).

Table 13: Percentage of individuals reporting problems on the EQ-5D descriptive system; the mean VAS by study sites

Usual Pain/dis­ Anxiety/ EQ-5D Mobility Self-care VAS activities comfort depress. N % % % % % Total 321 30.8 6.9 17.8 50.2 39.9 69.0 Southern Italy

.m Gallicianô 22 32 18 68 "t: " 36 ^ % - Castelmezz. 39 13 8 5 56 36 73

SE Spain Salobre 51 39 - 12 55 26 67

Viveros 71 18 11 14 27 3 67

Crete (Greece) Crete A 64 55 6 34 70 67 65

Crete B 74 26 7 18 43 65 72 Chapter 3 Socio-nutritional study 114

A quarter of the responders (27 %) perceived their own health as excellent or very good, 40 % stated their health as good and the remaining 33 % of household members reported their health as fair or poor (see Figure 23).

How would you rate your health during the past four weeks? 60

□ Excellent I Very good □ Good I Fair I Poor

I I

Galliciano Castelmez. Salobre Viveros Crete A Crete B Total

Figure 23: Percentage of responders rating their health on a scale from excellent to poor (Italy, Galliciano [N = 22]; Italy, Castelmezzano [N = 39]; Spain, Salobre [N = 51]; Spain, Viveros [N = 71]; Greece, Crete A [N = 64]; Greece, Crete B [N = 73]; all sites [N =320]).

In all the three countries, rheumatism and arthritis were the most diagnosed or treated diseases in the past year (see Table 14) in all three countries and high blood pressure/ hypertension was the second most reported diseases. Chapter 3 Socio-nutritional study 115

Table 14: Diseases diagnosed/ treated in the past year in the three countries (gender specific)

Diseases Italy [%1 Spain [%] Greece [%] TOTAL diagnosed/treated in Male Female Male Female Male Female [%1 the past year N = 18 N= 38 N = 48 N= 74 N = 56 N= 82 Rheumatism or 41 36 33 45 29 56 40.1 arthritis High blood pressure/ 41 35 38 20 27 37 33.0 hypertension Angina or heart . 11 21 20 16 20 14.7 problems Asthma/bronchitis/ 18 5 23 12 20 7 14.2 respiratory problem Other health problems 10 5 2 11 13 27 11.2 reported Gastric or intestinal 3 6 14 7 24 9.0 problems Renal/kidney 6 11 8 19 7 8.5 problems

Diabetes - 11 4 10 6 13 7.3

Diagnosis of high blood pressure or hypertension in women differs significantly in the three areas (chi square test p = 0.064). Only 20 % of the female responders in Spain reported high blood pressure or hypertension, which is significantly (p = 0.023) less then in Greece with 37 %. Heart problems or Angina varied from 11 % of the interviewees in

Italy to 20 % in Spain, and 20 % in Greece, with no significant differences in men and women. Chapter 3 Socio-nutritional study 116

Furthermore, no significant differences were reported for cardiovascular problems when comparing low and high consumers of WFPs. Only the diagnosis of diabetes, rheumatism and gastric problems was significantly lower in the “low consumer group”

(p=0.036) compared to high consumers.

3.4.7 Questionnaire surveys: Methodological problems for field-based ethno-

pharmacologists

The following section is part of a joint publication by S. Edwards, S. Nebel and M.

Heinrich (Edwards et al. 2005).

One of the favoured means of gathering quantitative data in the health and social sciences, including ethnopharmacology and medical ethnobotany, is by use of questionnaires. However, while it has been recognised that there is a need for culturally appropriate ways to ask questions in these surveys, little has been written about the potential difficulties of using questionnaires themselves in some cultural contexts.

The socio-nutritional questionnaire on the consumption of traditional food plants and its relation to social, economic and anthropological factors was developed for use in rural areas of the Mediterranean. In the Graecanic area 22 members of eight households were interviewed in 2003. As the community is very small, most of the housewives of the randomly sampled households had already shared their ethnobotanical knowledge about

WFPs in unstructured interviews the year before (2002). So, the question: “Why, haven 7

/ told you this before? ” was very common, and made people at first a bit reluctant to continue the interview. However, after encouraging and explaining the purpose of the Chapter 3 Socio-nutritional study 117 interview, most of the people were happy to continue. In general, the sample size in this project is very small and statistical analysis might be problematic.

Furthermore, as the interview was on several sheets of paper, it looked very official to the people and increased their doubts. “Can our knowledge be just in figures? ” asked one of the interviewees. As at first the interview was conducted in Italian by the author and not the local dialect, the sound of the official language of Italy reinforced the impression of being something official to the people. Later the interview was conducted with the help of a local person from the village, to bring it closer to the people and reassure them, that the information gathered is entirely for this project and has nothing to do with the authorities. This, however, raised the question of the influence of a local translator present at the interview. Field-based ethnopharmacological and ethnobotanical research is very dependent on, and sensitive to, micropolitics, i.e. researcher-collaborator relations. The act of formal surveying through use of questionnaires can alter the status quo by creating a perceived imbalance in power between researcher and interviewee.

From the perspective of a questionnaire respondent, the formal survey makes them an

“object of study”, rather than a colleague on an equal footing.

Another methodological problem was the length of the interview. In this research, the interview had several parts, including as well a Food Frequency Questionnaire (FFQ). In the FFQ people were asked to give the frequency and average amount of different foods that they have consumed over the last 12 months. As mentioned by Shahar et al. (2003), a major dilemma of an FFQ is the length of the questionnaire. Shorter FFQ’s would save Chapter 3 Socio-nutritional study 118 time and energy for the interviewee, but an abbreviated FFQ may be less sensitive to traditional foods, and thus be less accurate.

There are advantages and disadvantages to both qualitative and quantitative approaches.

One advantage of qualitative research includes “flexibility”, thus allowing researchers to be innovative, although critics say that this may result in anecdotal or unstructured data, a failure to test hypotheses, and a tendency to bias. However, quantitative survey methods are not necessarily value free, and often manipulate data according to preconceptions. Extrapolation from sample surveys may ignore possible disparities that exist in the “real world”, and how respondents answer questions in formal contexts may differ from responses given in a more relaxed interview. Dependence solely on quantitative methods omits to consider the social and cultural construction of

“variables”, which need to be comprehended to obtain a complete and valid picture

(Silverman 2001). Scientific data that are based on repeatable experiments or random sampling is not necessarily superior to data generated qualitatively and are based on social realities. Science, after all, is a process of inquiry, whichever method is used. In the area of ethnopharmacology we need to focus on developing novel methods of inquiry, and on implementing the knowledge ethnopharmacologists gain to benefit the traditional custodians of this knowledge. Chapter 3 Socio-nutritional study 119

3.5 Conclusion

The selected rural Mediterranean areas and specifically Galliciano are an example of rapidly changing communities where local traditions compete with “modem” ways of life. The present study showed how the traditional consumption of wild food plant species is still strongly embedded in local cuisines. The habit of collecting and cooking edible WFPs is still alive among the older generation. But already today, a lot of the traditional knowledge regarding food use is no longer actively used by the younger generation. The fact, that WFPs are appreciated in mral communities can be ascribed to many factors: the perceived healthiness, taste appreciation as well as “sense of local/cultural identity”. Younger people, who are not able to identify, and to gather many of the culturally important WFPs of the Graecanic area, still eat wild food plants collected and prepared by their parents, as a supplementary source of vegetables. No difference regarding the frequency and amount of consumption was reported between the different age groups in the mral village in Southern Italy.

People who traditionally gather wild greens knew about the additional health beneficial properties of wild gathered greens. Sometimes they deliberately choose wild species available to them and seek desired pharmacological effects consuming traditional dishes including wild gathered plants. Hence, food may be medicine and vice versa medicine may be food.

In general, there is evidence that the diet in the Mediterranean countries has been affecting the health of the respective populations and the overall Mediterranean dietary Chapter 3 Socio-nutritional study 120 pattern is considered important for longevity. However, a major nutritional concern in the Mediterranean countries is the loss of the traditional Mediterranean dietary pattern.

Compared to the originally described Mediterranean diet, Italy shows a relatively poor compliance, meaning a trend to Westernisation of the dietary habits, while Greece and

Spain adhere closer to the Mediterranean dietary pattern. Reasons for this development might be found in the substantial socio-economic changes throughout all of Europe over the past 40 years.

The observed departure from the Mediterranean diet in the Mediterranean countries might be accompanied by the loss of its protective effects on health and, consequently, a rise in diet-related diseases in the future, such as cardiovascular diseases and cancer.

Although the nature of this epidemiological study does not allow causality to be inferred, the associations between “healthy” nutritional behaviours and the risk for cardiovascular disease observed suggest that certain lifestyle behaviours might place an individual at lower risk for cardiovascular disease. Elevated consumption of the neglected Mediterranean wild green plants and moderate-to-low consumption of meat and products, seem to be cardioprotective via the enhancement of the antioxidant potential. Whether the adoption of frequent wild green plant consumption by individuals will result in a lowering of the prevalence of CVD remains to be elucidated.

The quantitative nutritional aspects were assessed with a food frequency questionnaire.

This food consumption study was made at a representative period in the annual cycle, that is the spring season, when WFPs are available. Although the FFQ aims at including food eaten during the whole year, species that are available later in the year, such as Chapter 3 Socio-nutritional study 121 many fruits species, or mushrooms might therefore be underrepresented in the study.

Furthermore, the cross-national comparison might be problematic as it is known, that any dietary recommendations should be based on prevailing food patterns in a population in order to be relevant and suitable to the specific population (Gerber et al.

2000). Chapter 4 Antioxidant activity 122

4 Antioxidant activity of wild food plants

U

Signora Tota with a bunch of Lactuca viminea,

Galliciano 2002 Chapter 4 Antioxidant activity 123

4.1 Introduction

4.1.1 Health beneficial effects of the Mediterranean diet

In the last decade many pharmacological studies have focused on the prototypical

Mediterranean diet and its commonly used plant species and plant products, like garlic

(Kim et al. 2000; Baneijee et al. 2003; Rahman & Lowe 2006), olive oil (Visioli et al.

1998; Owen et al. 2000; Briante et al. 2002; Tuck & Hayball 2002), grapes and red wine

(Carluccio et al. 2003; Auger et al. 2004; Urquiaga & Leighton 2005; Monagas et al.

2006) or tomato (Re et al. 2002; Weisburger 2002), to gain a better understanding of the beneficial effects of the Mediterranean diet (de Lorgeril & Salen 2001; Galli & Visioli

2001).

However, the diversity and the role of WFPs in local nutrition and cuisine of the

Mediterranean has been neglected in pharmacological studies. So far, only very little is known about nutritional and pharmacological properties of WFPs that are consumed as vegetables in rural areas in the Mediterranean, as for example in the Graecanic communities in Calabria, Southern Italy. The consumption of these WFPs is highly relevant for health, as they often contain higher amounts of bioactive compounds than plants that have been under cultivation for many generations (Schneider 1982; Lionis et al. 1998; Trichopoulou et al. 2000; Stepp 2004). Already in the 1960-1970s different research groups showed that WFPs contain higher amounts of carotene (provitamin A) and vitamin C, compared to cultivated plants (Muskat & Keller 1968; Zennie &

Ogzewalla 1977). The nutritional potential of non-cultivated vegetables native to the

Mediterranean and the Near East, has been recognised as an important area of research Chapter 4 Antioxidant activity 124

(Guil Guerrero et al. 1998; Tarwadi & Agte 2003; Turan et at. 2003; Zeghichi et at.

2003; Nebel et al n.d.), but there is still a lack of phytochemical and pharmacological knowledge on WFPs, as only very few studies have been conducted in the last five years

(Trichopoulou et al. 2000; Pieroni et al. 2002a; Couladis et a l 2003; Simopoulos 2004;

Salvatore et al 2005).

This chapter provides a general overview of the outcome of the pharmacological evaluation of wild gathered Mediterranean food plants of the whole consortium (with the focus on aging-related, degenerative disease and cognitive decline), and in particular highlights the research that was conducted by the author in the framework of this LFN project.

4.1.2 Oxidative stress and natural antioxidants

In this part of the PhD-project, new sources for natural and nutritional antioxidants are explored, through primary screening o f WFPs collected in Southern Italy, South Eastern

Spain and Greece. Interest in the search for new natural antioxidants, such as polyphenols, has grown considerably over the past years because free radicals, reactive oxygen species (ROS) and oxidative stress have been shown to be linked to ageing related illnesses (ARIs) (Finkel & Holbrook 2000; Vaya & Aviram 2001; Perry et al.

2002) and a large number of other illnesses. Furthermore, it is known that antioxidants obtained through the diet have a significant impact on health (Trichopoulou et al 1999a;

Heim et al. 2002; Scalbert et a l 2005b; Stanner 2005; Visioli et a l 2005; Zem &

Fernandez 2005). Polyphenols have been repeatedly shown to attenuate the formation of oxidative stress markers in various cell-free, cell-based as well as in animal and, though Chapter 4 Antioxidant activity 125 to a lesser degree, human studies (Frémont et al. 1998; Kim et a l 2003; Prior 2003;

Rajendran et al 2004). Moreover, restriction of the use of synthetic antioxidants, such as butylated hydroxyanisole and butylated hydroxytoluene, due to their toxicity (Namiki

1990) has been an important incentive for research on novel natural antioxidants.

Free radicals are produced in the body as by-products of normal metabolism and as a result of oxidative stress, such as exposure to radiation or environmental pollutants

(Benzie & Strain 2005). Elevated levels of free radicals and reactive oxygen species

(ROS), lead to the occurrence of oxidative stress, a characteristic of most ageing related degenerative diseases (Butterfield et a l 2002; Molavi & Mehta 2004; Scalbert et al

2005b). Normally free radicals are neutralised by efficient systems in the body that include the antioxidant enzymes (superoxid dismutase, catalase, and glutathione peroxidase) and the nutrient-derived antioxidant molecules (vitamin E, vitamin C, carotenes, or polyphenols) (Halliwell & Gutteridge 1999). In healthy individuals, a delicate balance exists between free radicals and antioxidants. In some pathologic conditions such as diabetes, and in critically ill patients, oxidative stress causes the level of antioxidants to fall below normal (Sabu & Kuttan 2002; Maritim et a l 2003).

The pharmacological effect of polyphenols, a diverse class of natural products

(flavonoids and phenolic acids) of plant origin, are attributed to their antioxidant, anti­ inflammatory, antibacterial, as well as gene expression-modifying effects (Cao et a l

1997; Harbome & Williams 2000; Rice-Evans 2001; Depeint et a l 2002; Bode & Dong

2003; Scalbert et a l 2005a; Sies et a l 2005). Antioxidants can exercise their protective properties at different stages of the oxidation process and by different mechanisms Chapter 4 Antioxidant activity 126

(Halliwell et al 1995). Two main types of antioxidants are described: “primary” (chain breaking, free radical scavengers) and “secondary” or “preventive” antioxidants which may operate by variety of mechanisms, including deactivation of metals, inhibition of enzymes, scavenge oxygen, regeneration of “primary” antioxidants, etc. (Gordon 1990;

Aruoma 2003; Masella et a l 2005). The in vitro methods to analyse antioxidant activity are numerous. In this chapter two assays to determine antioxidant activity of plant extracts are described in detail.

The first assay used in this study measures the free radical scavenging capacity of a compound or extract {primary antioxidant). 1,1 -diphenyl-2-picrylhydrazil radical

(DPPH) is a molecule containing a stable free radical. In the presence of an antioxidant, which is able to donate an electron to DPPH, the violet/purple colour, which is typical to free DPPH radical (see Figure 24), decays and changes in absorbance at 517 nm can be analysed spectrophotometrically. This assay can provide information on the ability of a compound to donate a hydrogen atom, on the number of electrons a given molecule can donate, and on the mechanism of antioxidant action. Chapter 4 Antioxidant activity 127

N:H

AH

Na HO,

DPPH • (DPPH): H violet yellow

Figure 24: (DPPH):H is a product of the reaction between DPPH* and an antioxidant AH: (a stable free radical 2,2-diphenyl-1-picryl-hydrazyl DPPH*, CigHizNgOe, M= 394,33 g)

In addition to the DPPH assay, the antioxidant activity of the plant extracts was tested using an enzyme inhibitory assay^ as it is known, that some compounds can contribute to the decrease in oxidative stress via inhibition or activation of regulatory enzymes

(secondary antioxidant), as for example the xanthine oxidase (Hearse et a l 1986;

Koyamae^û/. 1999).

Xanthine oxidase (XO), a flavoprotein, catalyses the oxidation of hypoxanthine to xanthine which is further converted to uric acid (Figure 25). Upon oxidation of xanthine or hypoxanthine to uric acid, the dehydrogenase produces NADH, whereas the oxidase generates superoxid anions (O 2 ) (Stocker & Keaney 2004). In several pathological states (e.g. hepatitis, brain tumours, or postischemia tissue injury), XO serum levels are significantly increased and the reactive oxygen species ROS generated in the enzymatic Chapter 4 Antioxidant activity 128 process are involved in oxidative damage (Shieh et a l 2000; Borges et a l 2002). These reactive oxygen species (O 2 ") are also key players in chronic inflammatory conditions such as rheumatoid and other types of arthritis (McCord 1985; Klein et a l 1996;

Hagfors et a l 2003; Masseoud et a l 2005). Xanthine oxidase inhibitors are also used as antioxidant therapy for premature infants treated with hyperoxia, to improve the antioxidant status of lung tissues and fluids and to decrease the generation of superoxide radicals from the hypoxanthine/xanthine oxidase system (Stone 1999).

Xanthine Figure 25: Xanthine Oxidase (XO) O2 r generating superoxid anions (O 2 ”) and XDH XO hydrogen peroxide (H 2 O2) with the NADH < r 1k O2” and H2O2 formation of uric acid as the final product. Uric acid XDH = Xanthine dehydrogenase

In 1992 Cotelle and colleagues showed that the inhibition of superoxide anion production in the xanthine/xanthine oxidase system through flavonoids was as a result of both scavenger activity and inhibition of the enzyme. They concluded, that natural compounds, which are able to inhibit the XO as well as to scavenge superoxide anions, may be useful as protecting agents against cellular injury during reperfusion of ischemic tissues (Cotelle et a l 1996). Thus, it is important to link the measurement of the scavenging activity of extracts (e.g. in the DPPH assay) with the inhibition of the XO

(Cotelle et al 1992). Chapter 4 Antioxidant activity 129

4.2 Aims and Objectives

This project, based at the interface of food and medicine (health food, functional food and food supplements), helped to gain a better understanding of wild food plant species and their potential role as antioxidants. It was a significant intermediary step in the search for plant extracts with antioxidant activity, which finally lead to the identification of plant extracts, which may serve as a lead in the development of new health food supplements. The objective was to:

• Evaluate the primary in vitro antioxidant activity of wild food plant species of

the Mediterranean (Southern Italy, Spain and Greece) with established

antioxidant assays (Free radical scavenging activity with DPPH assay and

enzyme inhibition in the XO-System).

4.3 Materials and methods

In our research group (including the author, Sabine Nebel, and two Erasmus students,

Sandra Maier and Lydia Tomantschger) two spectrophotometrical methods were used to determine antioxidant activity of the selected plant extracts. Firstly the free radical scavenging activity (FRSA) was measured in the 1,1 -diphenyl-2-picrylhydrazil radical

(DPPH) assay, and secondly the inhibition of the enzyme xanthine oxidase (XO), which was evaluated in the xanthine/xanthine oxidase superoxide scavenging system.

4.3.1 Collection of plant material

Bulk samples of a total of 127 wild food plant species were collected in South Eastern

Spain, Greece (Crete) and Southern Italy. The samples from Southern Italy (N = 25) were collected in Calabria by the author (Sabine Nebel) and in Lucania by Dr. Andrea Chapter 4 Antioxidant activity 130

Pieroni in the spring 2002. The samples from Spain (N = 66) and Greece (N = 36) were collected by other members of the “Local Food - Nutraceutical” (LFN) consortium.

Only the edible plant parts of commonly used WFPs (leaves, seed, flowering tops, stems, petals, leaf stalks, or young aerial parts) were collected and dried at room temperature (see Figure 26). About one kilogram of dried plant material was collected in order to have an adequate amount of plant material from one season to investigate in the laboratory.

Figure 26: Drying edible parts of WFPs collected in spring 2002 on an old bed frame, at room temperature in Galliciano, Southern Italy

4.3.2 Preparation of crude plant extract

Extraction of plants form Southern Italy took place at the School of Pharmacy in London and Spanish and Greek samples were extracted in Murcia (Spain) by Rafael Llorach. A standardised extraction protocol was used at both locations. Dried and powdered plant Chapter 4 Antioxidant activity 131 material (60g) was extracted in 500ml ethanol (90 %) under reflux for 30 minutes. After the extract had cooled down to room temperature, it was filtered through filter paper

Whatman No.l, and the volume of the extract was concentrated under vacuum (rotary evaporator, 40°C). Finally, the extract was freeze-dried (-50°C) and stored at -20°C.

Extracts were re-dissolved in ethanol or DMSO prior to usage in the individual assays.

4.3.3 Antioxidant assay: measurement of free radical scavenging activity (FRSA) in

the 1,1 -diphenyl-2-picrylhydrazil radical (DPPH) assay

The free radical scavenging activity (FRSA) assay was carried out according to the method used by Pieroni et al. (2002). 10 mg of the crude extract were dissolved in 5 ml of ethanol: water (4:1) and 1.5 ml of this solution was added to 1.5 ml of DPPH (1,1- diphenyl-2-picrylhydrazil) 0.1 mM dissolved in DMSO. Absorbance was read at 517 nm after 2 min. The FRSA was calculated as [ 1 -(Ai-Aj)ZAc]* 100, where Aj is the absorbance of 1.5 ml of the crude extract solution mixed with an equal volume of the

DPPH solution; Aj the absorbance of 1.5 ml of the crude extract solution mixed with an equal volume of DMSO, and Ac the absorbance of a blank prepared mixing 1.5 ml of the

DPPH with an equal volume of ethanol: water (4:1). Quercetin (a frequently occurring and well studied fiavonoid) and trolox (water-soluble Vitamin E) were used as standard antioxidant. In addition, an extract of bulbs of Leopoldia comosa (Liliaceae) and an extract of the aerial parts of Rhodiola rosea (Crassulaceae), widely sold in health food stores because of its reported radical scavenging activity (Furmanova et al. 1998), were used as controls. The experiments were performed in triplicates. Chapter 4 Antioxidant activity 132

4.3.4 Enzyme inhibition assay: Inhibition of xanthine oxidase

The xanthine/xanthine oxidase superoxide scavenging system used in our study evaluates the inhibition of the enzyme XO, or the scavenging activity of the sample against superoxide free-radical anions O 2 " (Gordon 1990). The inhibition of the XO was measured using a modified spectrophotometrical method base on Noro et al. (1983),

Owen and Johns (1999) and Cimanga et al. (2001). The activity of XO was measured by means of the amount of uric acid produced by the system. The plant extract (15 mg) was dissolved in 1.2 ml of ethanol and 1.8 ml of phosphate buffer (pH 7.5). The solutions were centrifuged, and 0.2 ml of the supernatant was added to the assay mixtures [2.7 ml of phosphate buffer, 2.0 ml of xanthine (0.15 mM). Finally, the reaction was started by adding 0.1 ml xanthine oxidase from buttermilk to the assay mixture (0.39 U/ml dissolved in phosphate buffer). At this concentration of ethanol (less than 2 % v/v) XO activity was little affected (see also Noro, 1983). The assay mixture was incubated at

37°C for 60 minutes. The uric acid production was measured at room temperature at

295 nm against a blank solution (assay mixture, without plant extract). The XO inhibition was expressed as percent inhibition of XO, calculated as (1-B/A) * 100, where

A is the change in absorbance of the assay without the plant extract (absorbance with

XO - absorbance without XO), and B is the change in absorbance of the assay with the plant extract (absorbance with XO - absorbance without XO). Positive control was allopurinol 2.5 mM, a well known XO inhibitor (Klein et a l 1996). The experiments were performed in triplicates. Chapter 4 Antioxidant activity 133

4.3.5 Further assays part of the LFN analysis

Members of the LFN consortium conducted a set of primary assays focusing mainly on aging-related, degenerative diseases, such as CVD, cancer, diabetes type II, and cognitive decline (The Local Food-Nutraceuticals Consortium 2005). An overview is presented in Table 15.

Table 15: Assays conducted by the LFN partners (The Local Food-Nutraceuticals Consortium 2005)

Antioxidant assays: • Measurement of hypochlorous acid-induced oxyhaemoglobin bleaching (OxyHb). • Measurement of lipid peroxidation in mouse brain tissue (MDA). • Measurement of plant extracts’ protective ability against hydrogen peroxide- induced DNA damage (Comet). Enzyme inhibitory assays: • Inhibition of acetylcholine esterase assay (AChE). • Inhibition of myeloperoxidase-catalysed guaiacol oxidation (G-OH). Anti-inflammatory (Griess-) assay [including cytotoxicity assessment (Viability)] : • Inhibition of cytokine-induced cell activation (Nitrite). • Cytotoxicity assessment (Viability). Anti-proliferation assay: • Determination of endothelial cell proliferation (BrdU). Anti-diabetic assay: • PPARy binding assay (PPARy). Mood disorder-related assay: • Serotonin re-uptake inhibition (hSERT). Determination of polyphenol (PP) content present in plant extracts Chapter 4 Antioxidant activity 134

4.3.6 Phytochemistry of Reseda alba L.

The aim of this side project, conducted by Master student Henrik Nilsson from Uppsala

University, Sweden under the supervision of Sabine Nebel, Marco Leonti and Michael

Heinrich, was to isolate novel compounds from Reseda alba L. (Resedaceae) with pharmacological potential. The focus was primarily set on isolating flavonoid-type compounds, since many flavonoids have pharmacological potential (Cook & Samman

1996; Kartnig & Buschmann 2005).

The dried plant material (890 g dried young leaves) ground and extracted with dichloromethane and methanol (cold percolation) yielded in 18 g dichloromethane extract and 225 g methanol extract respectively. After liquid-liquid partitioning the dissolved methanol extract (solvent water) with ethyl acetate, the ethyl acetate extract was fractionated by vacuum liquid chromatography (VLC). Flavonoid-type compounds were isolated using preparative thin layer chromatography (PTLC) and the purity of the samples was verified with thin layer chromatography (TLC) and *H-NMR (Nuclear

Magnetic Resonance). The structure was elucidated using NMR and Mass Spectrometry

(MS).

4.4 Results and Discussion

The results obtained in all the above-mentioned assays/systems, their linking and comparison can provide important information on the assessment of antioxidant properties of the plant extract. Thus it appears that it is important, not only to present the data obtained by the author, but to include all results obtained in the primary screening by the whole LFN - Consortium. Assays, which were conducted as part of the PhD Chapter 4 Antioxidant activity 135 project in hand, and plant species that were collected in Southern Italy, are highlighted in grey.

In total, 127 plants belonging to 42 different plant families have been collected in

Greece (N = 36), Spain (N = 6 6 ), and Italy (N = 22) (see Appendix 8.3). All plants presented are spermatophytes except three, which are mushrooms {Agaricus campestris p# 1, Lactarius deliciosus p# 120 and Terfezia arenaria p# 124). The plant parts used as food in the local communities were extracted using a standardized protocol. The consortium applied a set of primary assays focusing mainly on aging-related, degenerative diseases, such as cardio vascular disease (CVD), cancer, diabetes type II, and cognitive decline. Also, several EC 50 values have been determined for standard antioxidants to allow a comparison of the results with data from the literature: E C 5 0 f o r quercetin = 9.3 pM (MDA), 6.9 pM (GOH), 2.25 pM (Comet); E C 5 0 for luteolin = 77 pM (OxyHb); E C 5 0 for trolox (water-soluble Vitamin E) = 120 pM (MDA), 118 pM

(GOH), 380 pM (OxyHb). All screening data obtained in the various cell-free and cell- based assays are represented by symbols (Table 16) highlighting a high (#), medium (I), or low (O) biological activity in the specific assay and as compared to the controls of the extract. Chapter 4 Antioxidant activity 136

Table 16: Definition of activity range (% of control) for the performed bioassays.

Highlighted in grey: assays conducted by the author and Erasmus students at the School o f Pharmacy, UK.

Activity Concentration Number Assay (mg extract/ High ( • ) Medium (1) Low (O) ml)

Antioxidant assays

1 DPPH # ^50% 30%-49% <30% 1.0

2 OxyHb >50% 25 % - 50 % <25% 0.2 3 MDA >75% 50 % - 75 % <50% 0.2 4 Comet >40% 40 % - 20 % <20% 0.01

Enzyme inhibition assays

5 XO 220% 10% -19% <10% 0.1

6 AChE >20% 10% - 19% <10% 0.1

7 G-OH >75% 50 % - 75 % <50% 0.2

Anti-inflammatory assay (including cytotoxicity assessment) 8 Nitrite <30% 30% -70% >70% 0.1 9 Viability >90% 75 % - 90 % <75% 0.1

Anti-proliferation assay 10 BrdU >50% 30%-50% <30% 0.1

Anti-diabetic assay 11 PPARy >75% 30%-75% <30% 0.1

Mood disorder-related assay 12 hSERT >75% 30% - 75 % <30% 0.1 Chapter 4 Antioxidant activity 137

Furthermore, a total activity score (TAS) was calculated for each extract in order to identify those plants with the most promising profile of biological effects. For this, following activity scores were assigned: • = 1,1 = 0.5, 0 = 0. The TAS calculation for an extract was accomplished by adding up the activity scores from the 12 bioassays

(Table 17).

Table 17: Summary of primary screening data (TAS = total activity score; n.d. = not determined: ct = cytotoxic). Highlighted in grey: vertical = assays conducted at the

School of Pharmacy by the author with Erasmus students; horizontal = plant samples from Southern Italy. Key to # see Appendix 8.3 (Number = Plant species).

>> .£ 00 Anti­ Antioxidant assays Enzyme inhibitory inflammatory/ h assays cytotocity i l l -11 l3 i II 1 < a X < w X CO £ o a "C o X o s u 9 2 c/2 < Q U < u CQ 2 JD H o > % Dm 1 O o o n.d. o O # n.d. n.d. ct e O 2 2 o o i n.d. o i o 1 1 O i o 2.5 3 o o o i o # o e •O o o 3.5 4 1 o o # o ct ct 1 1 2 5 o 1 1 O # o i o O e 1 o 4 6 o o 1 O # o e e e e o o 5 7 1 o i o o o o o 1 ct e o 2.5 8 o o • o o # e e # n.d. o o 5 9 o 1 o 1 # 1 e # o 1 e i 6.5 10 o 1 1 o o o i e 1 • e o 5 11 o e o o i 1 1 e i 1 o o 4.5 12 O' o o o ë o 1 e o e o o 2.5 13 o o e n.d. # e o ct n.d. e o 4 14 o 1 e O i e e i O o o 4.5 15 o o e o o e o ct ct e o 3 16 t o e o 0 i e e e n.d. o 1 5.5 17 o o 1 o è i 1 e o O o 1 4 18 t o e o # i • 1 e n.d. o o 5.5 19 1 o o o o o 1 o O # o 2.5 H LA LA LA LA LA LA LA LA LA -fi- 4i- -P- -f^ -f^ -p- -p- -Pk Lk» Lk) Lk» Lk) Lk» Lk» Lk» Lk) Lk» Lk» K) K) N) K) K) W N» NJ w K) # U) LA Lk) 0 0 ON LA ■Pk Lk» t o 0 0 '-J O n LA -Pk Lk» II) 0 0 ON LA K) O NO 0 0 -<1 è O NO o NO O 2 ■ o ?r Q

4Êk âÊk m m m m éÊtk DPPH #OO O o e e O OO o O o OOO OO O O e o OO O O o o r ' o 3 O O O O O O •O O O OO O O O OxyHb 5‘ e o o o o o o o o o o o o o o C S- e e • o # e e e o o m o o e •• o e e • e e e o o o # e MDA

3 3 3 3 Cornet o o o o o o o e o o o o O o o o p - o o o o o o o o p . o • o o p - o o 9^ o

3 m , P XO o o o o o o # o e o o m o o O O o e e e o o o # o o o o o o o CL o P- V 3 âÊk AChE o o o o e o o O p . o o o o o o o o o o o e o o e o e e e ##e o o O o o O e # e e o e o o o e e o o o • e G-OH

3 3 Viability e o o o o •# e o e e e e # e e ##p . o e e e e m o • pk e

3 3 m Nitrite o o e o o o m o o o o o e o m e p . o # o e p- e

3 3 3 3 3 3 3 m o m m BrdU o o Cu o p - o ÇL o o o o e o o o O e o e o p . o e o p . p- e

m, m PRARy o o o o o e e o e # e % o o • o e o # e eo o e o O o o o O # O#o

e o Oo o o o o o o Oo o o o o o o e o o o o e o e e o e O m o o O o hSert

UJ -fi- L»J -p- 4 ^ OJ Lk) K) -p^ LA Lk» O n 4ik -Pk -f^ -pk Lk) NJ K) LA NO LA Lk) Lk) ON ON -pk Lk» Lk» -Pk Lk» K) -pk Lk» LA TAS LA LA LA LA LA CLA LA LA LA LA LA LA LA LA LA LA LA LA LA U) oo H o \o \o\o vo O nD 00 00 OO 00 00 oo OO OO 00 --4-4 -4-4 -4 -4 ~4'-o--4 ONONON ON ONONONONLA # D) =r o\ LA U>w 8 Ooo ONLAS LUK) o nO 00 ONLA3! LUw o NOoo -4s; LA£LUK) ONO a; eu fC "S # -Pu o o mko O o OOOo OOO#OOOOOOo Om o m o o o O o o m mm m DPPH o 5 m, P OxyHb 5’ o • #o o o o # o puOo o o o o o O o o o o O o o o OOO c o. P p o o e e e e e e e # o o p- p. o e e • o o o étkOOo o o o o o o o e o o MDA

p P p p p p p o o o o o o o o o e p- eu p. o e o o o o o o o o o o o p. p. p. o o p. Cornet

P o o o o o o o o o o o o o o o o o o o o o o XO o e e #o #o e o pu o o e p p o o ##o o o p- o o o p. o m o o o o o o o AChE o e e e o e e e e o pp. e o o o e o o o o o o o o o o o o o o o o G-OH p # e e e e e e e e# ep- e e • o e e e m#e e o e # • e eo • Viability o m o o o o # p-p o o o o e o o o o o o o o e o o o Nitrite p p p p p p p p p BrdU o m, o. e a. a . p- p. o o p. o pu o o p- # o o m o o o o o e o p- e o e O o o o O• o o o o o p.P o o o o e o o e m o o o o o e e e o o o o o o PRARy

P o o o e o O o o e o Pu o o o o o o O o o o o o m #o o o hSert

LA p LU LA o NO NO 4 ^ LU o ■fiu ON LA LA LA LA LA LA 4u LU 4u LA NO LU - LU 4 u LU LA - NO LA TAS - 4^ LA NJ 4^ LA LA LA LA LA LA LA LA LA w \o Chapter 4 Antioxidant activity 140

Table 17 continued

x':i X < a X 5 u Xfi 4fc >> Q s q z ■ok, 0> < & o s PÛ Xi H o U 6 > z pL. 98 Cl o o o % ct O m • i2.5 99 o O o O o 1 o e i 1 e 1 4 100 o o o n.d. o o o o ct e e o 2 101 o 1 o O o o o o ct n.d. o o 0.5 102 1 • e O o 1 # e 1 n.d. e o 6.5 103 o o o o o o o e O O o o 1 104 i o o o o e o e O e 1 1 4.5 105 o o o n.d. e 1 e # 1 o n.d. n.d. 4 106 o « o O o e # o 1 e # O 5 107 o o o o o i o e 1 o O o 2 108 1 i e o o # o o o e o 4.5 109 o o e o o i # e o n.d. o o 3.5 110 e 1 e o o i e e o 1 o o 5.5 111 o o e n.d. o 1 1 n.d. n.d. 1 o 1 3 112 1 • e O o o • O ct n.d. o o 3.5 113 o n.d. 1 o o i e 1 1 1 e 1 5 114 o 1 e o o i o 1 O 1 i 1 4 115 e o o o o o o e o O o o 2 116 o o 1 o o o i e i n.d. i o 3 117 e o o o o o o o o O o i 1.5 118 1 o o o o o o o o O o o 0.5 119 • 1 e o 1 o # e i o o o 5.5 120 o o o o o i o e 1 o o o 2 121 e o 1 o o o o e o o o o 2.5 122 not determined 123 O 1 o o o e o e e n.d. i o 4 124 O o o o o o o i i O o o 1 125 o o o o o o o e i n.d. o i 2 126 # i # o e • e e o n.d. o o 6.5 127 1 o # o < 1 e o o 1 o o 4

4.4.1 Antioxidant activity

Several extracts (N = 13; 10 %) showed high activity in at least two antioxidant assays, and none of the tested extracts showed high activity in all four assays (DPPH; OxyHb,

MDA and Comet). On the other hand, 16 extracts (13 %) showed low activity in all four Chapter 4 Antioxidant activity 141 assays. The samples showing high or low activity are from eleven to 13 different plant families, respectively. Consequently, this distribution does not allow for the identification of particular plant families with an above average number of highly active plant species. Furthermore, a substantial number of extracts (i.e. 36 %) showed high activity in terms of cellular protection from lipid peroxidation, whereas only 3 % of all samples were able to substantially attenuate HiOi-induced DNA damage.

4.4.2 Enzyme inhibition activity

Only two of the 127 screened extracts showed high activity in all three enzyme inhibition tests (xanthine oxidase inhibition (XO), inhibition of acetylcholine esterase

(AChE), and inhibition of myeloperoxidase-catalysed guaiacol oxidation (G-OH)).

These active plants are Berberis vulgaris (leaves and tender stems; p# 51) originating from Spain and Vitis vinifera (leaves; p# 126) from Greece. On the other hand, 27 extracts (21 %) demonstrated low effects in all three tests.

Interestingly, Berberis vulgaris (#51, Berberidaceae) and Vitis vinifera (#126, Vitaceae) extracts showed not only high activity in the xanthine/xanthine oxidase system, but also in two antioxidant assays (DPPH and MDA). Also Reichardia picroides (# 38,

Asteraceae) from Italy, Thymus piperella (#95, Lamiaceae) and Vitis vinifera from Spain

(young shoots; # 127, Vitaceae) showed high activity in the XO assay and medium activity in the DPPH assay. As highlighted in the introduction, the inhibition of

“superoxide anion - production” through flavonoids in the xanthine/xanthine oxidase system might be the result of both scavenger activity and inhibition of the enzyme.

Cotelle et al. (1992; 1996) showed the importance of structure-activity relationship. Chapter 4 Antioxidant activity 142 since hydroxy-flavones exhibited interesting antioxidant properties expressed either by their capacity to scavenge free radicals (DPPH or MDA) or to inhibit XO. Furthermore, it is known, that the number of phenolic hydroxyl groups, the extent and nature of the substitutions, and their specific position on the ring structures influences whether flavonoids function as modulators of enzyme activity or as antioxidant agents (Rice-

Evans 2004). These effects (scavenging activity and enzyme inhibition) may be useful in the treatment of pathologies in which free radical oxidation plays a fundamental role

(Berger 2005; McCord & Edeas 2005).

4.4.3 Results of further assays part of the LFN analysis

In the following a short summary of the results conducted by other members of the LFN consortium, with assays focusing mainly on aging-related, degenerative diseases, such as

CVD, cancer, diabetes type II, and cognitive decline (The Local Food-Nutraceuticals

Consortium 2005).

Anti-inflammatory activity

The MTT assay was performed to identify false positive results due to decreased cell viability or disturbed metabolism. Almost 10 % of the plant extracts had a cytotoxic/pro- apoptotic effect (assessed by the mainly mitochondria-dependent reduction of the MTT dye) in MBE cells. Cytokine-induced inflammatory response was efficiently suppressed by 12 of the non-toxic extracts. The amount of PPs present in these samples scattered between 0 and 367 mg/g extract, indicating no correlation between PP concentration and the observed anti-inflammatory activity. Chapter 4 Antioxidant activity 143

Anti-proliferation activity

The extracts’ effect on cell proliferation has been investigated in HMEC-1 by the BrdU incorporation assay. High inhibitory effects on cell proliferation have been found with

17 extracts belonging to 8 different plant families. The PP content of the positively scored extracts ranged from 11 to 285 mg/g extract, again suggesting that the detected activities are not necessarily linked to the polyphenol content of the respective plant extracts. The latter observation is supported by the data obtained for the samples from

Papaver rhoeas. Whereas both extracts prepared from plants coming from Greece (p#

104) and Italy (p# 106) affect BrdU incorporation by more than 50 % (i.e. show high activity), the PP content differs by more than 100 % (i.e. 120 mg/g versus 286 mg/g, respectively).

Anti-diabetic activity

The modulation of PPARy binding activity has been suggested as a promising target for the prevention and treatment of diabetes type II. A total number of 34 extracts has been identified to significantly affect PPARy binding and confirmed to induce a luciferase reporter gene under the control of a PPARy responsive element. These extracts belong to

13 different plant families and the PP content varied between 10 and 629 mg/g extract.

Interestingly, 4 of the 5 extracts prepared from plants belonging to the Liliaceae showed high or medium activity, whereas 5 of the 6 samples belonging to the Lamiaceae demonstrated only low modulatory effects on PPARy binding activity. Chapter 4 Antioxidant activity 144

Mood-disorder modulating activity

Less than 8 % (i.e. a total of 10) of the extracts affected serotonin re-uptake by more than 75 %, i.e. showed high activity. The PP content of these samples, belonging to 5 different plant families, ranged from 40 to 449 mg/g extract. Of note, both Cynara cardunculus samples, collected in Italy (p# 29) and Spain (p# 30), showed a high activity whereas their PP content differed by more than 100 % (449 mg/g versus

214 mg/g extract, respectively).

Total activity score (TAS)

To summarise, a number of plant species with very promising pharmacological profiles have been identified based on a total activity score (TAS) > 6, i.e. Berberis vulgaris

(TAS: 9; p# 51; Berberidaceae), Reichardia picroides (TAS: 7.5 and 6.5; p# 37 and 38;

Asteraceae), Scandix australis (TAS: 6.5; p# 9; Apiaceae), Satureja montana (TAS: 6; p# 94; Lamiaceae), Thymus piperella (TAS: 7; p# 95; Lamiaceae), Lythrum salicaria

(TAS: 6.5; p# 102; Lythraceae), and Vitis vinifera (TAS: 6.5; p# 126; leaves, Vitaceae).

There is only a low correlation between the total polyphenols content (PP) and the observed biological activity. Extracts showing relatively high activity despite a low PP content, such as Thymus piperella (TAS 7; PP 119 mg/g; # 95; Lamiaceae), Vitis vinifera (TAS 6.5; PP 98.3 mg/g; # 126; leaves; Vitaceae), Onopordum macracanthum

(TAS 6; PP 89 mg/g; # 35; Asteraceae) or Mantisalca salmantica (TAS 6, PP

118.8 mg/g; # 34; Asteraceae), are of considerable interest for subsequent studies (see dotted box in Figure 27). Interestingly the chemically very diversified Asteraceae are the main source of medicinal plant species used by traditional societies in a variety of Chapter 4 Antioxidant activity 145 applications in the northern hemisphere (Moerman et al. 1999; Leonti et al. 2003a;

Stepp 2004).

The following graph showing the correlation between the polyphenol content PP of the wild gathered food plants and the Total Activity Score (TAS), has to be read carefully, as the TAS is only an artificial score, summarising the whole range of assays.

700

600

500 & §: 400 c I 300

= 0.1555 1 200 S. 100

0 0 1 2 3 4 5 6 7 8 9 10 Total Activity Score (TAS)

Figure 27: Graph showing the low correlation (R^ = 0.16) between the Total Activity Score (TAS) and the polyphenol content (PP) of all plant samples (N = 127). Box highlights plants with relatively low polyphenols content and a high TAS.

The data for the PP content suggest that substantial amounts of non-phenolic constituents, such as bioactive terpenes or alkaloids, fatty acids or vitamins, are present in the extracts. This fact provides interesting lead for further nutritional-pharmacological studies. Chapter 4 Antioxidant activity 146

4.4.4 Phytochemistry of Reseda alba L.

The phytochemistry of Reseda alba is practically unknown. Only one compound, 2- hydroxy-2-methyl-propyl-glucosinolate, had previously been isolated (Gmelin & Kjaer

1970). Therefore our approach to the phytochemical investigation was fairly general. In total five fiavonoid glycosides were isolated from the methanol extract of Reseda alba.

One of the isolated fiavonoid (number 4) was identified as kaempferol-3,7-0-a-L- dirhamnoside (see Figure 28).

3" 2" 4" OH

Figure 28: kaempferol-3,7-0-a-L-dirhamnoside (kaempferitrin)

de Sousa et al. (2004) did research on the antioxidant potential of the pure compound kaempferol-3,7-O-a-L-dirhamnoside and they found, that the compound showed a very high reactivity in the DPPH assay, inhibited myeloperoxidase and decreased lipid peroxidation. Chapter 4 Antioxidant activity 147

4.5 Conclusion

There has been a growing interest in the value of polyphenols among researchers and food manufacturers. This is mainly because of their antioxidant properties, their abundance in our diet, and their probable role in the prevention of various diseases associated with oxidative stress, such as cancer, cardiovascular disease, and neurodegeneration (Scalbert & Williamson 2000; Bagchi & Preuss 2005). In recent studies, polyphenols have been shown to attenuate the formation of oxidative stress markers in various cell-free, cell-based as well as in animal studies (Frémont et a l 1998;

Kim et al 2003; Prior 2003; Rajendran et al 2004). Surprisingly, in this study none of the primary assays revealed a clear correlation between the observed biological activity and the polyphenols (PP) content, suggesting that the amount of PPs is not necessarily a predictive indicator for the potential antioxidant activity. This observation, however, is in contrast to recent studies showing good correlations between total PPs and focusing specifically on the total antioxidant activity (Leontowicz et a l 2002; Moyer et a l 2002;

Chinnici et al 2004; Loots et a l 2006).

The ethanol-based extraction procedure used in the present study aimed to optimise the amount of lipophilic bioactive compounds in the plant extracts. The author and the members of the consortium, however, were aware that highly lipophilic compounds were not extracted with this procedure. Also, the data for the PP content suggested that substantial amounts of non-phenolic constituents, such as bioactive terpenes and alkaloids, fatty acids or vitamins, were present in the extracts. Hence, since we used

“crude” extracts, the low correlation between the total PP content and the observed Chapter 4 Antioxidant activity 148 biological activities in fact provides interesting leads for further nutritional- pharmacological studies. Therefore, extracts showing high activity despite a low PP content are of considerable interest for subsequent studies.

Only a few studies investigated the biological activities of such wild gathered or semi­ cultivated plants (Trichopoulou et a l 2000; Grande et a l 2004; Schaffer et al 2004;

Simopoulos 2004). Species like Reichardia picroides, Scandix australis or Lythrum salicaria are largely unknown to the broader medical and nutritional community.

Although the primary screening has only relied on in vitro assays, the detected broad activity profile, covering antioxidant, enzyme inhibition, anti-inflammatory, anti­ proliferation, anti-diabetic as well as mood disorder-attenuating effects, make these plants very promising candidates for more detailed pharmacological in vitro as well as in vivo studies. In order to better understand the antioxidant activity of these WFPs, more phytochemical investigations are needed. The causal link between antioxidant administration and disease prevention or between lack of antioxidants and disease occurrence remains to be established, but the prudent public health advice continues to be to consume a variety of plant food. Chapter 5 Conclusion 149

5 Conclusion

The Graecanic villages are examples of rapidly changing communities where local traditions compete with “modem” ways of life. The present study shows how the consumption of wild food plant species is embedded in the local culture, providing a strong link between local people and their natural environment. However, recent changes in lifestyle result in the risk of losing this valuable local knowledge about WFPs and with it a healthier diet. Already today, a lot of the local knowledge regarding food use is no longer actively used by the younger generation and is subject to many outside influences and changes. This study has shown, that the practice of gathering and cooking of a diversity of edible wild plants is still alive, especially among the older generation.

Although WFPs have often been regarded as an emergency source of food in times of famine, they are still appreciated today. This can be attributed to several factors: the given “sense of local/cultural identity”, taste appreciation, as well as the perceived healthiness. Younger people, still eat ta chàrta (wild gathered greens) gathered by their parents, as a supplementary source of vegetables in spring, though they are no longer able to identify many of the culturally important wild greens of the area. In 2005, a handbook on the use of WFPs in the Graecanic area was published and distributed among the inhabitants of the area and at local schools to re-valorise such local food traditions (Nebel 2005).

The cultural importance of WFPs is also shown in the transmission of plant names in this traditional, oral knowledge system and the continuous use of WFPs as food. The fact that most plant names of the culturally important wild food plant species of the Chapter 5 Conclusion 150

Graecanic area have cognates with the same plant species in Greece indicates the historical importance of plant use. Etymologically these cognates have their origins in the Ancient Greek language and accordingly Ancient Greek culture. These findings make it highly likely that the continuous use of WFP species as vegetables, condiment or salads and the co-transmission of knowledge and practice has occurred since the time

Magna Graecia flourished in Southern Italy or at least since Byzantine times. The use of the historical-linguistic method facilitates the better understanding of the development of plant knowledge in cultures relying on oral transmission of knowledge. The further potential of using such an approach has to be explored and similar comparative analyses for other regions are needed to elaborate this method.

Compared to the originally described Mediterranean diet, the data from Italy showed a relatively poor compliance in the socio-nutritional study, showing a trend to

Westernisation of dietary habits, while the regions in Greece and Spain adhered closer to the Mediterranean dietary pattern. This socio-nutritional study emphasised the importance of wild foods in traditional diets, and showed that most of the gathered

WFPs were perceived to be healthy by the local people. The diversity of plant resources used has played an essential role for enabling populations to meet their nutritional, health and socio-cultural needs. Biodiversity equates with dietary diversity which equates with health. The convergence of medicinal and culinary practices was also shown in the Graecanic area. Several wild gathered food plants were consumed as food because of the attributed health beneficial effects or had a multifunctional use as food as well as medicine. Chapter 5 Conclusion 151

The biological activity of such wild gathered food plants has only been investigated in a

few studies in recent years. However, the consumption of these WFPs is highly relevant

for health, as they often contain higher amounts of bioactive compounds than plants that

have been under cultivations for many generations. The evaluation of the antioxidant

activity and other biological activity of WFPs used in the Mediterranean, showed

promising results for several plants (young leaves of Reichardia picroides, leaves and

tender stems of Scandix australis, and Berberis vulgaris, or aerial parts of Thymus piperella and Lythrum salicaria). Until now, several of these active plant species were

largely unknown to the wider medical and nutritional community, for example

Reichardia picroides. Although the primary screening has relied on in vitro assays, the

detected activity profile, covering antioxidant, enzyme inhibition, anti-inflammatory,

anti-proliferation, as well as anti-diabetic effects, make these plants very promising

candidates for more detailed pharmacological in vitro as well as in vivo studies.

Furthermore, more phytochemical investigations of WFPs are needed in order to better

understand the antioxidant activity.

This detailed assessment of local food habits in the Mediterranean sheds light on a

largely overlooked aspect of Mediterranean diets, and furthermore helps not only to

preserve but also to disseminate local knowledge regarding the nutritional suitability of

wild gathered fruits and vegetables. This study is based on a few selected regions in

Southern Europe and many other variants of Mediterranean diets, from such diverse

countries as Portugal, France, Albania, Cyprus, Turkey, the Arabic Mediterranean

countries, Lebanon, and Israel, remain undocumented and unstudied from an

ethnobotanical and pharmacological-nutritional perspective. Chapter 6 References 152

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complementary and alternative medicine among British undergraduate pharmacy

students. PWS Pharmacy World & Science (online 11 May 2006) Chapters Appendix 188

8 Appendix

8.1 Ethnobotanical questionnaire (in Italian)...... 189

8.2 Socio-nutritional questionnaire (in Italian)...... 193

8.3 List of wild food plants used in the primary screening

(Antioxidant activity) ...... 208

8.4 CD “ta chorta: Piante commestibili tradizionali di Galliciano’*...... 213 Chapter 8 Appendix 189

8.1 Ethnobotanical questionnaire (in Italian)

LOCAL FOOD - NUTRACEUTICALS Questionario

Numéro progressivo questionario: _____ Numéro progressivo pianta: _____

1. La pianta

Nome locale (in grecanico): ______(Numéro o codice di riferimento erbario: ______)

2. Dove si raccoglie

La pianta è: □ coltivata □ selvatica (spontanea) □ semi-coltivata? Se coltivata, area approssimativamente coltivata per famiglia: ______

Se non coltivata, qual è la diGusione di questa pianta in natura? □ molto rara □ rara □ frequente □ molto frequente

Questa pianta si trova al mercato: □ si □ no 3. Come si raccoglie

Quale parte è utilizzata: ______Quando viene raccolta? ______

Chi la raccoglie: □ donne □ bambini □ uomini □ anziani □ tutti e due

Quale strumento è usato per la raccolta? Chapter 8 Appendix 190

4. Uso in cucina

Come si mangia questa pianta: □ sola □ assieme ad altre piante (erbe/verdure) Se si, quali?______

Frequenza dell'uso di questa specie: □ più di una volta la settimana □ una volta la settimana □ una volta al mese □ qualche volta durante I'anno □ una volta all'anno o più raramente

Quando questa pianta viene consumata, quante volte si mangia came? □ più di una volta la settimana □ una volta la settimana □ una volta al mese □ qualche volta durante I'anno □ una volta all'anno o più raramente

Questa pianta era mangiata una volta durante la carestia (o in tempo di guerra)? □ si □ no

Cosa si fa per prepararla (prima di cucinarla)? Si lava? Si lascia nell'acqua? ______

Come si cucina?

Nome locale di questo piatto:_

5. Percezione delle sensazioni

Ha un sapore particolare: □ si □ no Chapter 8 Appendix 191

Se si, come descriverebbe questo sapore? (come si dice in grecanico?) □ acido □ amaro □ dolce □ salato □ astringente □ altro:______

Ha questa pianta un odore tipico: □ si □ no Se si, come descriverebbe questo odore? □ di fiori □ di legno □ di erba □ altro:

Quali persone NON dovrebbero mangiare questa pianta?_ Quali persone dovrebbe mangiarla preferibilmente? ____

Questa pianta fa bene alia salute? □ si, fa bene □ fa molto bene per la salute □ non lo so □ no, non fa bene □ altro:______

Se questa pianta fa bene alia salute, per quali malattie in particolare?

Se questa pianta NON fa bene alia salute, perché?

6. Altre informazioni Usi medicinal!:

Usi veterinari:

Usi nei lavorio agricoli (ad esempio per fabbricare attrezzi):

Usi magici/rituali:_

Leggende o storie che si raccontano su questa pianta:_ Chapter 8 Appendix 192

7. Dettagli sull'informatore

Nome ed indirizzo:

Sesso: □ F Eta:

□ M

Scuola: Lavoro (attuale e passato): ______Luogo dell'intervista (casa, piazza, strada):

Data:

8. Eventuale richieste dell'informatore □ Richiesta di omissione del nome (in base alle leggi vigenti sulla privacy) □ Richiesta di co-autorship nell'articolo Se si, con quale nome e cognome: ______Chapter 8 Appendix 193

8.2 Socio-nutritional questionnaire (in Italian)

EUROPEAN LOCAL FOODS PROJECT CONSORTIUM OFFICE Centre of Pharmacognosy and Phytotherapy Research The School of Pharmacy University of London 29/39 Brunswick Square , London, WCIN lAX

WP2: LOCALLY PRODUCED FOOD PRODUCTS: Interviews with householders

Notes to interviewer

Standard type: Directions to the interviewer Bold: To be spoken out loud to respondent Italics: Prompts to be used when needed

Interview no. Household code: Country code ....

Area code: .. Interviewer: D ate:...... / ...... /.

Prima di cominciare I’intervista, vorrei spiegarvi brevemente le ragioni di questo studio.

II proposito di questo studio e quello di analizzare Puso di prodotti alimentari vegetali usati localmente da diverse comunita in differenti regioni europee. Per saperne di pin, stiamo per questo intervistando persone in case selezionate casualmente.

In questa intervista, vorrei porvi domande sulPuso di prodotti alimentari, per sapere quale contributo questi apportano alia vostra dieta, come voi pensate che questi influenzino la vostra salute, e come e perchn voi consumate e scegliete cibi differenti.

Vi chiedero anche alcune informazioni sulla voi stessi, la vostra salute in generate, i vostri consumi alimentari quotidiani, sia a voi che ad altre persone della famiglia.

Assicurarsi che il formulario relative al consenso sia stato compilato. Chapter 8 Appendix 194

Sezione 1. Circa gli aliment: vegetali che voi consumate a casa

1. Avete voi la responsabilita principale per:

Decidere che cibi vengono consumât! nella casa? Si...... 1 No ...... 2 Responsabilita divise ...... 3 (per favore fomire dettagli)

Preparare i pasti? Si...... 1 No ...... 2 Responsabilita divise ...... 3 (per favore fomire dettagli)

2. Ci sono alcuni erbe spontanee prodotte o raccolte localmente che sono usate in questa area. a. Quanto spesso consumate [name of plant] quando e stagione? la maggior parte del giorni ...... 1 una volta alia settimana ...... 2 da una volta alia settimana ad una volta al mese ...... 3 raramente ...... 4 mai ...... 5 b. Ha [name of plant] qualche effetto utile/benefico alla/sulla vostra salute? Yes ...... 1 No ...... 2 Non lo so ...... 7 Non risposta ...... 9

Se si, per favore descrivete questi effetti con maggiori dettagli: (enter full response in table) Ha altri effetti? (enter full response in table) c. Ci sono rischi per la salute o possibili effetti negative associate con questo alimento? Yes ...... 1 No ...... 2 Non lo so ...... 7 Non risposta ...... 9

Se si, quali? (enter full response in table) Chapter 8 Appendix 195

Prompts which can be used to gather full information on this question: Pensate che ci siano effetti sulla vostra salute generale? Pensate ci siano effetti sulla vostra longevita? Pensate ci siano effetti su particolari condizioni o sintomi e, se si, quali?

Table 1: responses to question 2

Plant Name of plant Frequency of Perceived helpful/ Perceived risks/ code consumption beneficial effects on bad effects in season health 50 gattinaria {Reseda alba)

51 pricadida (Lactuca viminea)

12 gaddazida {Reichardia picroides)

13 masaro {Foeniculum vulgare)

14 rigano {Origanum heracleoticum)

3. Ci sono altre erbe spontanee coltivate o raccolte che consumate nella vostra famiglia? Si 1 No (go to q4) 2 Quali sono?

Obtain a list and enter into table 2. Chapter 8 Appendix 196

For each plant listed ask questions a, b, and c as in question 2. Table 2: responses to question 3

Plant Name of plant Frequency of Perceived helpful/ Perceived risks/ code consumption beneficial effects bad effects in season on health

4. Quanto sono important! 1 seguenti fattori nell'influenzare la vostre scelte alimentari? Per ognuno indicare se e molto importante, importante, ma solo qualche volta, non importante.

Non Importante Molto importante qualche volta importante 01 Tempo e convenienza nell'acquisto o 1 2 3 nella raccolta 02 Difficolta/facilita di preparazione 1 2 3 03 Disponibilita di prodotti diversi 1 2 3 04 Possibilita di conservazione a casa 1 2 3 05 Costo 1 2 3 06 Per mantenere una dieta sana/stare in 1 2 3 salute 07 Alimento che preferisco 1 2 3 08 Preferenza del sapore da parte della 1 2 3 famiglia e degli amici 09 Altro (specificare) 1 2 3 Chapter 8 Appendix 197

Quali sono i due fattori che voi considerate i pin importanti nelle scelte alimentari?

Enter code (01-09) from above table ......

Quali i due fattori meno importanti? Enter code (01-09) from above table ......

Sezione 2. Altri aspetti della vostra dieta e stile di vita

5a. Quale delle seguenti affermazioni a vostro riguardo e vera? Sono un fumatore (almeno una sigaretta, sigaro, pipata ogni giomo) 1 (go to 5b) Fumavo, ma adesso non sono pin un fumatore (cioe: ho smesso di fumare almeno mesi fa) (go to 6) 2 Non ho mai fumato (go to 6) 3

5b. Approssimativamente quante sigarette/sigari/pipate fumate di solito al giorno?

Enter actual or approximate number ...... not applicable ... .998

6. Fate lavori fîsici?

Si No

Se si, per favore potete descrivere cosa fate?

Descrizione attivita Quante volte nelle Ore in media in ultime due ogni occasione settimane 1

2

3 Chapter 8 Appendix 198

Fate esercizio fisico per passatempo?

Si ...... 1 No 2

Se si, potete per favore descrivere cosa fate?

Descrizione attivita Quante volte nelle Ore in media in ultime due ogni occasione settimane 1

2

3

Fo o d Freq uency Q uestionnaire

Media di consume durante gli ultimi 12 mesi

6+ 4-6 2-3 una 5-6 2-4 una pin 1-3 volte volte volte volta volte volte volta rara A LIM EN TO E volte al al al al alia alia alia ment QAUNTITA al giorn giorn giorn giorn setti setti setti e o mese 0 0 0 0 mana mana mana mai

LATTICINI

LATTE

intero (di mucca, di capra o di pecora)(l tazza=240 ml.) scremato (di mucca, di capra o di pecora) (grasso 0 % -l,5 %, 1 tazza=240 ml.)

YOGHURT

intero,incluse yogurt fatto in casa (1 tazza= 220m l.) scremato (grasso 0 % -l,5 %, 1 tazza=220ml.) Chapter 8 Appendix 199

a latte intero (formaggio prodotto localmente, pecorino, caprino, misto, parmigiano, mozzarella a latte intero (30 gram mi o 1 vaschetta)) a latte scremato (grasso 0 %-1,5 %, 30 grammi o 1 vaschetta))

CEREAL!

pane bianco (1 fetta=30 grammi) pane intégrale (1 fetta=30 grammi)

PASTA/RISO

spaghetti, pasta corta (1/2 tazza)

riso (1/2 tazza)

FRUTTA;:: / •-

mela (1 frutto o 1/2 tazza di succo)

pera (1 frutto 0 1/2 tazza di succo)

arancia (1 frutto o 1/2 tazza di succo)

pesca (1 frutto o 1/2 tazza di succo)

uva ( 1 tazza)

m andarino (1 frutto)

melone (1 fetta)

anguria o cocomero (1 fetta) Chapter 8 Appendix 2 0 0

ciliege (1 tazza=10 frutti)

fragole(l tazza=8 frutti)

banana (1 frutto)

ogni altro frutto (100 gram mi)

VERDURE E LEGUMI \ ^ 1 . 7 ' * . . pomodori (1 pom odoro di taglia m edia o 1 tazza di pomodori crudi) cetrioii (1 tazza crudi) spinaci (1/2 tazza cucinati, o 1 tazza crudi) nielan/ane (1/2 tazza cucinate)

zucchine (1/2 tazza cucinate) broccoli e cavolfiore (1/2 tazza cucinati) lattuga e radicchio (1/2 tazza cucinati, o 1 tazza crudi) ogni altra verdura (1/2 tazza cucinata, o 1 tazza cruda) piseili, lenticchie, fagiolini, fagioli (1/2 tazza cucinati) ■' V u v ' VERDURE SPONTANEE < ‘ ^ . . gattinaria {Reseda alba, 1/2 tazza cucinata) pricadida {Lactuca viminea, 1/2 tazza cucinata) gaddazida {Reichardia picroides, 1/2 tazza cucinata) masaro {Foenicidiini vulgare spp./?//?er/mw, 1/2 tazza cucinato) Chapter 8 Appendix 2 0 1

rigano {Origanum heracleoticiim) ogni altra verdura spontanea o miscele di queste (1/2 tazza cucinate)

CARNE

tacchino e polio (90- 120 gram mi cucinato, ad esempio un petto di polio arrostito)

GARNI ROSSE ' ■; . -V. if' manzo (90-120 grammi cucinato, anche com e m acinato)) maiale (90-120 grammi cucinato, anche com e m acinato) agnello e capretto (90-120 grammi, anche macinato)

PESCE

tonno (100-120 gram m i) sardine(100-120 grammi, fritte, arrostite, etc.) ogni altro tipo di pesce (100-120 gram m i)

G RASSI ED OLII

Che tipo di olio e/o grasso usate per cucinare? • olio di oliva • strutto • burro • olio di semi di girsaole • margarina • olio di semi di mais • altro tipo (per favore specificare) Chapter 8 Appendix 202

Che tipo di olio e/o grasso usate per condire I'insalata? olio di oliva strutto burro olio di semi di girasole maionese margarina sen ape creme a base di latte altri condimenti (per favore specificare)

Che tipo di olio e/o grasso usate sul pesce o sulla carne? olio di oliva strutto burro olio di semi di girasole maionese margarina senape creme a base di latte altri condimenti (per favore specifîcare)

Sezione 3» Voi e la vostra salute

Vorrei ora farvi alcune domande sulla vostra salute. Per favore indicate quali delle seguenti affermazioni descrivono meglio il vostro stato di salute oggi, segnando con una crocetta una sola casella di ciascun gruppo.

9. Capacita di movimento Non ho difficolta nel camminare n...l Ho qualche difficolta nel camminare 1 1 .. .2 Sono costretto/a a letto □... 3

10. Cura della persona Non ho difficolta nel prendermi cura di me stesso [ I... 1 Ho qualche difficolta nel lavarmi o vestirmi n .. .2 Non sono in grado di lavarmi o vestirmi □...3

11. Attivita abituali (per es. lavoro, studio, lavori domestici, attivita familiari a di svago) Non ho difficolta nello svolgimento delle attivita abituali Cl... 1 Ho qualche difficolta nello svolgimento delle attivita abituali 171 ...2 Non sono in grado di svolgere le mie attivita abituali 171 ...3 Chapter 8 Appendix 203

12. Dolore o fastidio Non provo alcun dolore o fastidio n...i Provo dolore o fastidio moderati n...2 Provo estremo dolore o fastidio II...3

13. Ansia o depressione Non sono ansioso o depresso 1:1...I Sono moderatamente ansioso o depresso 0...2 Sono estremamente ansioso o depresso IJ...3 Chapter 8 Appendix 204

14. Per aiutarvi ad esprimere il suo stato di salute attuale, Migliore abbiamo disegnato una scala graduata (simile ad un termometro) sulla quale il migliore stato di salute stato immaginabile contrassegnato dal numéro 100 ed 11 peggiore 100 dallo 0. Vorremmo che indicaste su questa scala quale e, secondo voi, il livello del suo stato di salute oggi, tracciando una linea dal riquadro sottostante fino al punto che corrisponde al vostro 9'LO stato attuale di salute.

0

II vostro stato di salute

CODE = NUMBER BETWEEN 000 AND 100 AS SPECIFIED BY RESPONSDENT ON THE SCALE 4j[0

PL C

0 Peggiore stato di salute immaginabile Chapter 8 Appendix 205

15. In generate, come definireste la vostra salute durante le ultime 4 settimane? Eccellente 1 Molto buona 2 Buona 3 Discreta 4 Poco buona 5

17. Ecco qui una lista di problemi di salute. Quali di questi vi sono stati diagnosticati o curati dal dottore durante lo scorso anno? Pressione alta o ipertensione S1...1 No...2 Angina or problemi cardiaci S1...1 No...2 Diabete S1...1 No...2 Reumatismi o artriti S1...1 No...2 Asma, bronchite o altri problemi respiratori Si... 1 No...2 Problemi gastrici o intestinali S1...1 No...2 Problemi renali S1...1 No...2 Altri (per favore elencarli) S1...1 No...2

Nessuno dei citati S1...1 No...2

18. Annotare il sesso dell'intervistato/a Maschio 1 Femmina 2

18. In quale anno siete nato/a?(enter year 19XX)

18b. Qual e la vostra eta in anni? .....anni

19. Come descrivereste la vostra situazione lavorativa? studente ...... 1 lavoro a tempo pieno ...... 2 lavoro a tempo parziale ...... 3 casalinga ...... 4 disoccupato/a ...... 5 pensionato/a ...... 6

19b. Qual e la vostra occupazione?

20. Qual e Poccupazione del capofamiglia?(se persona diversa da ql2) (please enter description)

not applicable ...... 8 Chapter 8 Appendix 206

21. Qual e il vostro livello di educazione piu elevato? livello elementare ...... 1 livello second ario (primo ciclo: circa dagli 11 ai 14 anni)...... 2 (secondo ciclo: circa dai 14 ai 18 anni)...... 3 livello universitario...... 4

21b. Quanti anni di scuola avete frequentato? ...... anni

22. Da quanti anni vivete in paese? ...... anni

Se venite da qualche altra zona,

dove vivevate prima? in un altro paese della Calabria .1 in un'altra regione italiana .2 in un'altra nazione .3 non applicabile .8

Se da questo paese,

vostra madre proveniva da questo paese .1 un altro paese della Calabria .2 un'altra nazione (per favore specificare) .3

vostro padre proveniva da questo paese .1 un altro paese della Calabria .2 un'altra nazione (per favore specificare) .3

24. A quale gruppo etnico appartenete? Italiano ,11 Grecanico ,12 Altro (per favore sepcificare) Chapter 8 Appendix 207

Sezione 4. Altre persone della casa

24. Quante persone vivono nella casa, incluso/a voi? (enter number) ...... (mangiano in media almeno una volta al giomo ogni giomo e dormono qui. Se ci sono variazioni stagionali, includere persone che erano nella casa la settimana precedente)

Per ogni persona della casa, per favore ditemi: in quale relazione di parentela lei/lui e con voi (1 coniuge, 2 figlio, 3 figlia, 4 cognato, 5 cognata, 6 altro grado di parentela, 7 nessuna parentela) sua eta (anni) 1 .maschio/2.femmina da quanto tempo vivete qui? (anni)

Relazione Eta Sesso Da quanti anni vive qui 1

2

3

4

25. Ci sono altri membri della famiglia che hanno vissuto qui e che ora hanno traslocato? Per ognuno/a di essi, per favore ditemi: in quale relazione di parentela lei/lui e con voi (1 coniuge, 2 figlio, 3 figlia, 4 cognato, 5 cognata, 6 altro grado di parentela, 7 nessuna parentela) sua eta (anni) 1 .maschio/2.femmina da quanto tempo sono partiti? (anni)

Relazione Eta Sesso Da quanti anni vive qui 1

2

3

4

Please report the approximate length of the interview (minutes) Chapter 8 Appendix 208

8.3 List of wild food plants used in the primary screening (Antioxidant activity)

Overview table on the taxa of local Mediterranean food plants investigated in the “Local

Food - Nutraceutical” project, their origin(Spain = S, Italy = I, Greece = G), and content of polyphenols (PP = amount of polyphenols in mg/g ethanolie extract). Highlighted in

grey = plants samples collected in Southern Italy.

Part(s) used for # Scientific name Plant family c culinary purposes ‘5b c CL O CL 1 Agaricus campestris L. ex Fr. Agaricaceae Caps I 1&2 2 Aizoon hispaniciim L. Aizoaceae Aerial parts S 46.7 3 Amaranthiis cf. graecizams L. Amaranthaceae Leaves G 2 5 ^ 4 Apiiim nodiflonim L. Apiaceae Aerial parts I 8&9 (Umbel liferae) 5 Dauciis carota L. Apiaceae Leaves G 243.7 6 Eiynghim campestre L. Apiaceae Sprouts S 55.1 7 Foeniculum vulgare Mill. Apiaceae Leaves G 178.87 8 Oenanthe pimpiuelloicles L. Apiaceae Leaves G 17.4 9 Scandix australis L. Apiaceae Aerial parts S 191.55 (leaves and tender stems) 10 Scamlix pecten-veneris L. Apiaceae Leaves G 130.7 11 Tordylium apuliim L. Apiaceae Leaves G 57.8 12 Phoenix dactylifera L. Arecaceae Tender leaves S 11.0 (Palmae) (very tender leaf rachis) 13 Achillea ageratum L. Asteraceae Aerial part S 214.7 (Compositae) 14 Anacyclus clavatus Pers. Asteraceae New shoots, s 54.5 tender leaves 15 Arctium minus Bemh. Asteraceae Tender leaves s 189.1 16 Artemisia alba Asso Asteraceae Aerial parts s 229.9 17 Carlina acaulis L. Asteraceae Flower I 59.1 receptacles 18 Chiliadenus glutinosus Fourr. Asteraceae Aerial parts s 253.3 19 Chondrillajuncea L. ^A st^ceae : Leaves I 11T8 4 20 Chrysanthemum coronarium Asteraceae Leaves G 235.1 L. 21 Chrysanthemum segetum L. Asteraceae Young leaves I 2 5 j 22 Cichorium intybus L. Asteraceae Leaves G 48.1 23 Cichorium intybus L. Asteraceae Leaves S 107.0 Chapter 8 Appendix 209

Part(s) used for 1 # Scientific name Plant family c culinary purposes '5b c a, O CL 24 CfcAofiuw L. # Asteraceae # Basal leaves I 64.9^11 25 Cichorium spinosiim L. Asteraceae Leaves G 40.1 26 Crepis commutata (Sprengel) Asteraceae Leaves G 115.4 W. Greuter 27 Crepis vesicaria L. Asteraceae Leaves G 148.5 28 Crepis vesicaria L. subsp. Asteraceae Tender leaves S 69.1 haenselerii 29 Cynara cardunculus L. subsp. Asteraceae Stems, flowers I 448.8 cardunculus and receptacles 30 Cynara cardunculus L. var. Asteraceae Rachis (leaf S 214.4 ferocissima Lowe rachis) 31 Helmintotheca echioides (L.) Asteraceae Leaves G 50.2 Holub. 32 Hypochoeris radicata L. Asteraceae Leaves I 215.9 33 Lactuca viminea L. Asteraceae Leaves I 210.2 34 Mantisalca salmantica L. Asteraceae Tender leaves S 118.8 Briq. & Cavill. 35 Onopordum macracanthum Asteraceae Tender leaves S 8 9 ^ Schousboe 36 Helmintotheca echioides (L.) Asteraceae Aerial parts I 242.1 J. Holub 37 Reichardia picroides Roth Asteraceae Leaves G 318.5 38 Reichardia picroides Roth Asteraceae Leaves I 335.1 39 Rhagadiolus stellatus (L.) Asteraceae Tender leaves S 24.4 Gaertner 40 Scolymiis hispanicus L. Asteraceae Rachis (tender S 110.8 leaf rachis) 41 Scorzonera cretica Will. Asteraceae Leaves G 151.3 42 Silyhum marianum (L.) Asteraceae Tender leaf rachis S 8&0 Gaertner 43 Sonchus asper L. Asteraceae Tender leaves s 108.3 44 Sonchus oleraceus L. Asteraceae Tender leaves s 75.2 45 Sonchus oleraceus L. Asteraceae Leaves G 122.0 46 Sonchus oleraceus L. . ^ A steraceae Leaves I 1572 j 47 Sonchus tenerrimus L. Asteraceae Tender leaves S 9 9 2 48 Urospermum dalechampii Asteraceae Tender leaves I 424.6 Scop. ex. F.W. Schmidt 49 Urospermum delechampii Asteraceae . Aerial parts I 8&6 Scop. ex. F.W. Schmidt Chapter 8 Appendix 210

ÛX) Part(s) used for # Scientific name Plant family ■S culinary purposes ‘C CL, o eu 50 Urospermum picroides (L.) F. Asteraceae Leaves G 245.9 Scop. ex. W. Schmidt. 51 Berheris vulgaris L. subsp. Berberidaceae Leaves and S 271.2 seroi O. Boles & Vigo tender stems 52 Berheris vulgaris L. subsp. Berberidaceae Fruits s 178.1 seroi 0. Bolos & Vigo 53 Anchusa officinalis L. Boraginaceae Leaves G 154.5 54 Borago officinalis L. Boraginaceae Leaves G 36.3 55 Borago officinalis L. Boraginaceae Leaf petioles S 5.3 56 Lithodora friiticosa (L.) Boraginaceae Aerial parts S 161.2 Griseb. 57 Symphytum hulbosiim Boraginaceae Leaves G 145.56 Schimp. 58 Brassica oleracea L. var. Brassicaceae Stems G 5&9 capitata (Cruciferae) 59 Eruca sativa Mill. Brassicaceae Leaves G 5&4 60 Eruca vesicaria Cav. Brassicaceae Leaves S 311.7 61 Erucaria hispanica Druce Brassicaceae Leaves G 123.2 62 Hirschfeldia incana (L.) Brassicaceae Leaves G 6T 6 Lagr.- Fossat 63 Hirschfeldia incana (L.) Brassicaceae Stems G 8T 6 Lagr.-Fossat 64 Moricandia arvensis (L.) DC. Brassicaceae Tender leaves S 23.1

65 Nasturtium officinale R. Br. Brassicaceae Leaves S 2 & 0 66 Raphanus raphanistrum L. Brassicaceae Leaves G 40.4 67 Sinapis alba L. Brassicaceae Leaves G 149.6 68 Sinapis arvensis L. Brassicaceae Aerial parts I 208.2 69 Sisymbrium crassifolium Cav. Brassicaceae Sprouts S 34.1 70 Opuntia ficus-indica (L.) Cactaceae Fruits S 9.2 Mill. 71 Capparis spinosa L. subsp. Capparidaceae Stems S 63.71 spinosa 72 Capparis spinosa L. subsp. Capparidaceae Flowers s 10.84 spinosa 73 Capparis spinosa L. subsp. Capparidaceae Fruits s 5Z 0 spinosa 74 Silene vulgaris (M oench) Caryophyllaceae Leaves I 162.2 Garke subsp. vulgaris ^ 75 Silene vulgaris (M oench) Caryophyllaceae Leaves s 119.4 Garke subsp. vulgaris Chapter 8 Appendix 2 1 1

Part(s) used for 1 # Scientific name Plant family c culinary purposes '5b •c Q- O a, 76 Beta vulgaris L. subsp. cicla Chenopodiaceae Tender leaves S 185.8 (L.) Arcangeli 77 Cheuopodiiim bonus-henricus Chenopodiaceae Leaves G 59.3 L. 78 Merendera montana Lange Colchicaceae Bulbs S 206.7 (Liliaceae) 79 Fallopia convolvulus (L.) A. Convulvulaceae Leaves S 118.2 Love 80 Bry’onia dioica Jacq. Cucurbitaceae Young shoots s 49.7 81 Cucurbit a pepo L. Cucurbitaceae Leaves G 157.6 82 Cyperus esculentus L. Cyperaceae Tubers s 22.5 83 Arbutus iinedo L. Ericaceae Fruits s 37.6 84 Lotus edulis L. Fabaceae, s.str, Young fruits I n.d. (Leguminosae) 85 Lupinus albus L. Fabaceae, s.str. Seeds s 40.4 86 Vicia faba L. Fabaceae, s.str. Leaves G 221.9 87 Vicia faba L. Fabaceae, s.str. Tender unripe s 76.3 fruits 88 Quercus ilex L. subsp. Fagaceae Fruits s 102.5 Rotundifolia (Lam.) T. Morals (Desf.) Samp. 89 Globularia vulgaris L. Globulariaceae Leaves s 207.3 90 Juglans regia L. Juglandaceae Fruits s 366.7 91 Hissopus officinalis subsp. Lamiaceae Aerial parts s 131.0 pilifer (Labiatae) 92 Mentha aquatica L. Lamiaceae Aerial parts s 100.8 93 Origanum heracleoticiim L. Lamiaceae Flowering tops I 65.2 94 Satureja montana L. Lamiaceae Aerial parts s 339.4 95 Thymus piperella L. Lamiaceae Aerial parts s 119.4 96 Thymus pulegioides L. Lamiaceae Aerial parts I 435.1 97 Allium ampeloprasum L. Liliaceae, s.l. Bulb s 4.8 (Alliaceae) 98 Asparagus acutifolius L. Liliaceae, s.l. Young shoots I 116.4 (Asparagaceae) 99 Asparagus horridus L. Liliaceae, s.l. Young shoots s 6&8 (Asparagaceae) 100 Muscari spreitzehoferi Liliaceae, s.l. Bulbs G 24.3 (Heldr.) Vierh. (Hyacinthaceae) 101 Ornithogaliim narbonense L. Liliaceae, s.l. Bulbs G 32^ Chapter 8 Appendix 212

To Part(s) used for Tb # Scientific name Plant family _c culinary purposes '5b •C c u O CL, 102 Lythnim salicaria L. Lythraceae Aerial parts s 628.7 103 Ficus carica L. M oraceae Fruits s 1.3 104 Papaver rhoeas L. Papaveraceae Leaves G 119.7 105 Papaver rhoeas L. Papaveraceae Aerial parts I 34T * (leaves, tender stems and buds) " I 106 Papaver rhoeas L. Papaveraceae Aerial parts I 285.8 107 Roemeria hyhrida (L.) DC. Papaveraceae Tender leaves s 83.9 108 Pimis piriea L. Pinaceae Seeds s 197.2 109 Plantago sempervirens Plantaginaceae Aerial parts s 163.5 Crantz. 110 Rumex thyrsyflorus Fingerh. Polygonaceae Leaves G 61.9 111 Rumex piilcher L. Polygonaceae Leaves G 230.2 112 Lysimachia ephemerum L. Primulaceae Aerial parts S 384.5 113 Clematis vitalba L. Ranunculaceae Young shoots I 222.5 114 Reseda alba L. Resedaceae Aerial parts I 186.3 115 Ziziphusjujiiba Mill. Rhamnaceae Fruits s 44.4 116 Crataegus monogyna Jacq. Rosaceae Fruits s 70.4 117 Primus cerasifera L. c.v. Rosaceae Fruits s 79.3 '''mayeras'' 118 Pyrus rehderi c.v. “Per et as ” Rosaceae Fruits s 126.4 119 Rubus ulmifolius Scott Rosaceae Sprouts s 92.1 120 Lactarius deliciosus (L. ex Russulaceae Carpophore s 3&9 Fr.) S.F. Gray 121 Citrus limon L. c.v. “Verna” Rutaceae Fruits s 78.5 122 Veronica beccabunga L. Scrophulariaceae Aerial parts I n.d. 123 Solanum nigrum L. Solanaceae Leaves G 11.3 124 Terfezia arenaria (M oris) Terfeziaceae Carpophore s 1 3 ^ Trappe 125 Celtis australis L. Ulmaceae Fruits s 0 126 Vitis vinifera L. Vitaceae Leaves G 9 8 3 127 Vitis vinifera L. Vitaceae Tender shoots s 235.9 Chapter 8 Appendix 213

8.4 CD “ta chorta: Plante commestibili tradizionali di Galliclano”

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