Fraochan - non timber forest products in Scotland

Fraochan
“That Which Grows
Amongst the Heather”
DEVELOPING A SUSTAINABLE
BLAEBERRY INDUSTRY FOR
SCOTLAND
Heather J Griggs
Dissertation, BSc (Hons)
Herbal Medicine
Napier University, Edinburgh
May 2007
Acknowledgements:
Richard Constanduros (Highland Natural Products)
Douglas Hardie (Highland Natural Products)
Graham Strachan (Highland Council)
Fiona Sinclair (Project Blaeberry)
Sarah Allen (Highland Council)
Anne Thompson (Ella Drinks)
And
With grateful thanks to Liz Craigen without whose support and generosity this
research would have been impossible.
2
Dedicated to my dear friend and classmate,
Arthur Clibury.
You will be profoundly missed.
3
Contents
Summary of Work
1
Chapter 1.1: Vaccinium
2
myrtillus in European History
Chapter 1.2: Vaccinium
5
myrtillus in Scottish History
Chapter 2: Vaccinium
7
myrtillus (L.) Botany
Chapter 3.1.1: Chemical
11
Composition of Vaccinium
myrtillus
Chapter 3.1.2: Antioxidant
15
Profiles and Recent Research
Chapter 3.2: Biomedical
18
Research: Recent Trends and
New Horizons
Chapter 4.1: The Role of
22
Vaccinium myrtillus in the Wild
Food Industry
Chapter 4.2: Vaccinium
26
myrtillus Crop Trials
Chapter 4.3: Harvesting and
30
Processing
Chapter 5: Discussion and
34
Conclusion
References
40
Appendix One:
51
Bibliography
Appendix Two:
65
Reproduction and Mycrorrhizal
Dependence
4
Appendix Three: Flavonoid
68
Anthocyanin Chemistry
Appendix Four: Blaeberry
72
Leaflets
5
Summary of Work
Background: Used for millennia worldwide as a food and medicinal plant, the
berries of Vaccinium myrtillus L., commonly known as bilberries, or blaeberries in
Scotland, have drawn increasing interest from the medical and health industries
since scientific studies were first performed on the extracts in the 1950s. More
recent studies on the antioxidant properties of bilberry anthocyanins have shown
potential for medical applications ranging from rheumatoid disorders to vascular
disease, cancer to diabetes. Whist the bulk of the world’s blaeberries are now
harvested in Scandinavia and Eastern Europe, recent investigations show that
France, Germany, Portugal, Russia, America, Canada, Japan, and Australia are
investigating sustainable sources of this valuable crop. Most Scottish fruit
producers have shunned any attempt to grow or harvest native blaeberries, while
Finnish companies harvest millions of tons per annum. Purpose and Methods:
This research will take a multi-disciplinary approach to the question of developing
a blaeberry industry in Scotland, covering such diverse areas of study as
ethnobotany, botany, biochemistry, medical research, funding and legislation,
non-timber forestry products, sustainability, industrial research, and harvest and
production methods already in place in other countries. Using a thorough
literature review, information on industrial processes, government reports, and
information gathered from those involved in recent blaeberry trials, this paper will
attempt to outline the possibilities and problems with industrial-scale harvesting
and processing of blaeberries in Scotland. Finally, this work will attempt to
provide a solid basis for future research into blaeberry production in Scotland and
the study of Scottish blaeberry antioxidants.
6
Chapter 1: Introduction
1.1 Vaccinium myrtillus in European History
Humans have been using Vaccinium species as food and medicinal plants
for many millennia. In Europe, wild cranberry (Vaccinium oxycoccus) residues
were found at a Bronze Age archaeological site in Denmark (Trehane 2004, 15),
and Vaccinium spp. pollen was found in the digestive tract of the Tyrolean
Iceman discovered in the Alps in 1991 (Dickson et al. 2000, 1845). Vaccinium
myrtillus (L.), widely known as bilberry, or blaeberry in Scotland, is the most
commonly found Vaccinium species in the wild in Europe (Trehane 2000, 18). In
the13th century, abbess and herbalist Hildegaard of Bingen (Figure One) claimed
in her herbals that blaeberry fruits induced
menstruation, and the 16th century German herbalist
Hieronymous Bock asserted that blaeberries could
be used to treat bladder stones, lung, and liver
disorders (Trehane 2004, 20). The first Amsterdam
Pharmacopoeia (1636) also includes an entry for
Fructus Myrtilli and a recipe for Oleum Myrtillorum
(Koning and Arnold 1961). Curiously, Morazzoni
and Bombardelli (1996, 4) claim that it was not
named in many European herbals before the 16th
century, but they do note, however, that the berries
were used traditionally in many European countries for diarrhoea, dysentery,
haemorrhoids, gastrointestinal inflammation, scurvy, and urinary complaints.
Decoctions of the leaves were also used internally for diabetes and externally for
inflammation of the eyes and mouth, infections, and burns (ibid.).
In Britain, blaeberries have also been in use for thousands of years.
Found on upland moors and in woods in areas as diverse as Somerset,
Derbyshire, Pembrokeshire, and throughout Scotland, the berries of Vaccinium
7
myrtillus are known by a variety of names across the British mainland (Figure
Two).
Common Name
Region/County
Blaeberry
Scotland, Cumbria, Lancastershire, Northumberland, Shropshire, Yorkshire
Blueberry
Cumbria, Yorkshire
Brylocks
Scotland
Hartberry
Dorset, Somerset
Hurtleberry
Devon, Somerset
Hurts
Cornwall, Devon, Gloucestershire, Hampshire, Surrey, Sussex, Pembrokeshire
Whortleberry
Somerset, Wiltshire
Whinberry/Wimberry
Cheshire, Derbyshire, Gloucestershire, Herefordshire, Shropshire
Figure Two: Common names for Vaccinium myrtillus in Britain
(After Trehane 2004)
Blaeberry remains have been found at diverse archaeological sites in
Britain including a Neolithic burial cairn in Sutherland (Dickson and Dickson
2000, 63) and near a Roman fort in Carlisle (ibid, 123). In British historical
literature, Culpepper makes mention of blaeberries in his famous herbal of 1653
(1981, 19):
It is a pity they are used no more in physic than they are. The black
bilberries are good in hot agues, and to cool the heat of the liver and
stomach; they do somewhat bind the belly, and stay vomitings and
loathings; the juice of the berries made in a syrup, or the pulp made into a
conserve with sugar, is good for the purposes aforesaid, as also for an old
cough, or an ulcer in the lungs, or other diseases therein.
Maud Grieve (1971), in whose early 20th century classic A Modern Herbal
blaeberry holds an entry, notes that the leaves can be used for urinary tract
disorders and for diabetes, whilst the fruits are good for diarrhoea and
‘discharges’. However, in all of the 19th century materia medicae (Murray 1825;
Duncan 1829; Craig 1879; and Leonard 1892) available for examination by the
author, blaeberries seem to have disappeared completely. C. Pierpont Johnson
(1862, 163), who includes a section on blaeberries in his mid-19th century ‘A
8
Treatise Upon the Principle Native Vegetables Capable of Application as Food,
Medicine, or in the Arts, and Manufactures’, comments that, at the time,
blaeberries were picked and consumed in large quantities in the rural areas of
Britain where they grew most abundantly, but were ‘less esteemed’, especially in
London, after the introduction of the cranberry from the Balkan states.
9
1.2 Vaccinium myrtillus in Scottish History
Being a country made up largely of heath land and open mixed forest, it
seems likely that Scotland would have always produced an abundance of
blaeberries for use by its animal and human populations. The Gaelic name for
Vaccinium myrtillus is fraochan from the word fraoch, or heather. It is listed in
MacBain’s Etymological Gaelic Dictionary ([online] MacBain1982) as meaning
‘heather-protector’, after its association with the top part of a walking shoe. Also
known as fraoch nan curra-bhiteag and fraochan caora-mhiteag (Comhairle nan
Eileen Siar 2007), fraochan can be loosely translated as ‘that which grows
amongst the heather’. Camilla and James Dickson’s Plants and People in
Ancient Scotland (2000) catalogues archaeological remains of blaeberry found
various sites around Scotland including a Neolithic site in Embo, Sutherland that
yielded two carbonized berries (Dickson and Dickson 2000, 63), a 12th century
site in Perth (ibid. 180), 13th century sites in Aberdeen and Paisley (ibid, 189,
196), and a 14th century site in Elgin, which contained a vessel with blaeberry
seeds in it (ibid, 190). Examination of the initial archaeological reports from the
Soutra Hospital Project (SHARP) excavations in the Scottish Borders (Moffatt et.
al. 1988; 1989) revealed no Vaccinium remains that could be strictly attributed to
human usage, but more recent excavations have revealed remains which
principle archaeologist Dr. Brian Moffatt attributes to usage by locals for
treatment of intestinal parasites (Vickers 2005).
Despite the ubiquitous presence of blaeberries in the Scottish landscape,
there are not as many historical or cultural references as one might expect in
Scottish literature (Sinclair 2000, 3). In her Millennium Award funded Blaeberry
Project (2000), Fiona Sinclair lists only a handful of references to blaeberries in
Scottish literature from the 15th through the 19th centuries. The Flora Celtica
database ([online] RGBE 2001), compiled by botanists at the Royal Botanic
Garden Edinburgh, also contains a small number of entries with reference to
10
historical blaeberry use in Scotland including uses as a dye, and for food and
medicine. In her book Healing Threads: Traditional Medicines of the Highlands
and Islands, Mary Beith (1995, 206) catalogues some common medicinal uses
for blaeberries including to ease dysentery, diarrhoea, and as a tea for dissolving
kidney stones. Allan and Hatfield (2004, 123) reiterate these uses, and add that
the berries were also used for cold symptoms and sore throats. Likewise,
Sinclair (2000, 4-5) quotes a number of historical sources from the 18th to the 20th
centuries that proclaimed the usefulness of Vaccinium myrtillus for ‘fluxes’, or
bleeding from the orifices, diarrhoea, and kidney stones. Perhaps the dearth of
historical references to blaeberry use can be partly attributed to the fact that they
were such a part of the landscape that their use was taken for granted (Sinclair
2000, 3). The author, however, suspects multiple forces at play, including the
economically, politically, socially, and linguistically fractured landscape of
Scotland. Although blaeberry jam was reportedly first imported to the court of
James V of Scotland in 1513 by the cooks of his French bride (Trehane 2004,
18), it seems more than likely that the peoples of Scotland have used blaeberries
in syrups and tinctures, honeys and preserves for at least as long as they have
been settled here.
11
Chapter Two: Vaccinium mytillus (L.) Botany
2.1 Botany and Ecology
The genus Vaccinium, belonging to the family Ericaceae, contains 450
species that are found across the globe. Of these, six can be found in the wild in
Europe. In his definitive book The Genus Vaccinium in North America (1988,
130), Vaccinium expert S.P. Vanderkloet identifies Europe’s native low-bush
blueberry, Vaccinium myrtillus (Linneus) as follows:
Shrubs (10) 17 – 45 (60) cm high, rhizomatous, forming open colonies.
Twigs green, conspicuously 3-angled, glabrous. Leaves broadly elliptical
or ornate, 7 – 11 mm wide, 19 – 27 mm long, green, laxly glandular
beneath; margin serrate. Calyx continuous with pedicel, green and
glabrous; lobes very small or absent; margin of the calyx tube merely
sinuate. Corolla globose, pink, cream, or greenish – white, 5 – 7 mm
wide, 3 – 5 mm long; lobes very small. Filaments glabrous, anther sacs
awned; pollen tetrads 34 – 38 µm in diameter. Berry purple, black, usually
not glaucous, 7 – 9 mm in diameter; nutlet approximately 1 mm long.
Chromosome number 2n = 24.
Figure Two
http://www.ag.uidaho.edu/sandpoint/images/Bilberry%20(Vaccinium%20myrtillus).jpg
12
Figure Three
http://www.treesforlife.org.uk/images/blaeberry2_200.jpg
Lang (1987, 148) provides a detailed description of the habitat of
Vaccinium myrtillus as it is found across the British Isles. For northern areas, he
explains that blaeberry is:
…abundant on heaths, moors, and oak, birch, and pinewoods on acid
soils throughout northwest Britain at altitudes up to 1200m. The dwarf
shrub zone composed of bilberry and cowberry, is one of the main
features on the mountains above the heather zone and below the open
summits, and it may form isolated island communities in sheltered, welldrained places on otherwise exposed bare tops especially where the rocks
break the wind. They are a characteristic feature of the Cairngorms above
1000m although they are absent from patches where the snow lives late in
the season.
He continues (1987, 149):
Bilberry can tolerate very dry conditions and extremely acid peat, often
growing with cowberry, and it can tolerate shade better than heather, so it
persists on wood margins. It probably reaches maximum growth and
reproductive performance in open pinewoods such as those in the Spey
Valley and it always fruits best with a little shelter.
Although blaeberry is able to grow at altitudes up to 1000m, it prefers drier, acid,
and shady environs, like those found in the open birch and pine woodlands of the
Caledonian Forest (Featherstone 2002) (See Figure Four).
13
Figure Four: Blaeberries growing in Clashwood, Ross and Cromarty
Taken by the author, July/August 2006
It likes to grow on the forest floor over old tree trunks and rocks and on some
moors and heaths it can be an indicator of woodlands having been there
previously (ibid.). Whilst blaeberry is plentiful in the north and west of Scotland, it
has declined over the past 300 years due to the destruction of Scotland’s native
woodlands and over-grazing by large-scale sheep husbandry (ibid.).
Vaccinium myrtillus plants are usually pollinated by bumblebees (Bombus
ssp.) or wasps, but may also self-pollinate to some degree (Featherstone 2002).
In the field of horticultural science, there are ongoing, international studies of
pollination by various insects within the Vaccinium genus, and these are easily
accessed via the internet from websites such as the International Society for
Horticultural Science Acta Horticulturae site (http://www.ishs.org/acta/index.htm).
The berries themselves develop soon after pollination in May (the author has just
seen the first berries of 2007 on May 1st) and usually ripen by the end of July or
14
early August. Vaccinium myrtillus, like some other fruiting trees and shrubs, are
subject to masting1 cycles of 3 to 4 years (Selas 2000, 424), a process that may
have some impact of blaeberry cultivation.
Reproduction methods are the subject of a good deal of research on
Vaccinium myrtillus and many of these studies address the matter of sexual
versus clonal reproduction. Sources (Featherstone 2002; Nuortila et. al. 2002;
Vanderkloet and Hill 1994) suggest that Vaccinium myrtillus does not have a
successful rate of sexual reproduction because seed germination can vary
considerably depending on the environment, the time in the season, and whether
the seeds have passed through the digestive system of a foraging bird or
mammal. A 1994 study of Canadian soil samples by Vanderkloet found that
seeds from this genus were underrepresented in the seed banks of soils where
they are found in North America (Vandekloet and Hill 1994, 56) even where the
plants were growing abundantly. He hypothesized that loss of seeds may be
taking place through dispersal by birds, between seed deposition and
establishment in the soil, through germination, or because of fungal
decomposition (ibid.). Contrary to earlier studies (as cited in Vanderkloet and Hill
1994, 57), a recently published study by Honkavaara et. al. (2007) found an
increase in rates of germination for Vaccinium myrtillus seeds when they were
passed through the digestive systems of thrushes (Turdus spp.). They found that
germination rates increased for ‘passed’ seeds except those from berries picked
very early in the season (Honkavaara et. al. 2007, 15). Variation in seed
germination rates also occurred between sample-years, leading them to
hypothesize that that there is potential variation in germination success within the
berry season itself, between crop years, and when seeds are eaten by birds or
foragers (ibid., 15 – 16). Clearly there is scope for continued research in this
area. (See Appendix Two for more information on blaeberry reproduction and
ecology).
1
In some plant species, individuals store up energy for two or more years, in
order to produce a large seed crop in one season. When all of the plants in a
population are synchronised to perform this mass seed production in the same
year, it is called masting (Selas 2000, 423).
15
Chapter Three: Biochemistry and Pharmacological Applications
3.1.1 Chemical Composition of Vaccinium myrtillus (L.)
Jean Bruneton (1999, 361 - 362) lists the chemical composition of bilberry
fruits and leaves in his comprehensive treatise on plant chemistry,
Pharmacognosy, Phytochemistry, Medicinal Plants:
[Blaeberries] are rich in water…, sugars…, and organic acids. Phenolic
acids, flavonoids (hyperin…,quercitrin), proanthocyanidins…,and
monomeric flavan-3-ols (catechin and epicatechin) have been identified.
The anthocyanin level in the fresh fruits is about 0.5%. These glycosides,
about fifteen of them, are C-3 O-glucosides, O-galactosides, and Oarabinosides of cyanidin, peonidin, delphinidin, malvidin, and petunidin.
The bilberry leaf contains phenolic acids, flavonoids…, and traces
of quinolizidine alkaloids… . It is rich in proanthcyanidins and catechin (up
to 10%) and in the 1950s, hydroquinone and arbutin were isolated. Later
on, it was not possible to characterize them again.
In fact, knowledge of Vaccinium myrtillus berry and plant chemistry has improved
much in the last two decades due to economic interest and advancing chemical
assay techniques. Morazzoni and Bombardelli (1996, 6) report the presence of
glucosides, galactosides and arabinosides of delphinidin, cyanidin, petunidin,
peonidin, and malvidin in a purified Vaccinium myrtillus berry extract called
Myrtocyan®, manufactured by Italian phytochemical giant Indena. They also cite
an earlier German study, which found 109 constituents in the berries including
aliphatic alcohols, aldehydes, ketones, terpene derivatives, and aromatic
compounds (Morazzoni and Bombardelli 1996, 5). According to various
European studies (as cited in Morazzoni and Bombardelli 1996, 5) the leaves
have been shown to contain quercetin and its glycosides, (+)-catechin, (-)epicatechin and their gallates, catechic tannins, iridoids, derivatives of cinnamic,
chlorogenic, and caffeic acids, and trace amounts of arbutin and hydroquinone
(ibid.).
16
Isolation and identification of flavonoid anthocyanins2 from blaeberry fruits
has been a subject of considerable research since the 1990s because they are
well known to be highly antioxidant. Blaeberries are also known to contain the
highest quantity of anthocyanins of all berry varieties, with a ratio of 30:36:13
parts for cyanidin, delphinidin, and malvidin 3-O-glycosides, respectively (Du et.
al. 2004, 60). Two papers from issue 49 of the 2001 Journal of Agricultural and
Food Chemistry examined the identification of anthocyanins in Vaccinium
myrtillus by two different methods of high-performance liquid chromatography
(HPLC)3, the most common method for identifying anthocyanins. Dugo et. al.
(2001) used a technique called electrospray ionization mass spectrometry (ESIMS) combined with the HPLC method to examine anthocyanin extracts from
various berries to elucidate differences between them. This technique confirmed
the identification of 14 anthocyanins in Vaccinium myrtillus including 3-Oarabinosides, 3-O-glucosides, and 3-O-galactosides of cyanidin, delphinidin,
peonidin, petunidin, and malvidin (Dugo et al. 2001, 3989), as reported by
Bruneton (1999) (See Figure Seven).
Figure Seven: Anthocyanins Identified in Blaeberry
delphinidin-3-galactoside
delphindin-3-glucoside
cyanidin-3-galactoside
delphidin-3-arabinoside
cyanidin-3-glucoside
cyanidin-3-arabinoside
petunidin-3-glucoside
petunidin-3-galactoside
peonidin-3-galactoside
petunidin-3-arabinoside
peonidin-3-glucoside
malvidin-3-galactoside
malvidin-3-glucoside
malvidin-3-arabinoside
After Dugo et. al (2001), 3990.
2
For a detailed explanation of flavonoid anthocyanin chemistry, see Appendix
Three.
3
High performance liquid chromatography (HPLC) is a commonly used
technique for separating, identifying, and purifying many chemical compounds.
See HPLC: A User’s Guide at http://www.pharm.uky.edu/ASRG/HPLC/hplcmytry.html.
17
This research is significant because it helps with the formation of a library of
anthocyanin fractions for production as commercial standards and with the
identification of anthocyanins in other matrices (ibid.).
Nyman and Kumpulainen (2001, 4183) used HPLC in combination with
diode array detection (DAD) to discern the anthocyanidin content of several
berries. They assert that the antioxidant behaviour of anthocyanins is related to
hydroxyl substitutions made on the B-ring, which leads to increased antioxidant
capacity. Thus methods for identifying the various types and quantities of
anthocyanidins present in berries may play a significant role in understanding the
benefits of these foods and the pharmacological agents that might be produced
from them. They make several important points in their discussion. Firstly, they
observe that the quality of anthocyanin standards available for purchase is poor
because they are unstable (Nyman and Kumpulainen 2001, 4185). This may
lead to possible biases in research due to selection and use of a single standard
reference glycoside for calibration, for instance (ibid.). Secondly, as mentioned
earlier, the structure of an anthocyanin is pH dependent and solvent systems for
samples may affect absorption characteristics (ibid.). With their HPLC-DAD
method, Nyman and Kumpulainen elicited an improvement in the sensitivity of
the identification of anthocyanidin peaks on a chromatograph and they
reconfirmed the method over 14 days to insure its repeatability without excess
variation. This kind of pure chemistry is imperative to finding better methods of
identifying and quantifying anthocyanins for use in other kinds of research.
A number of other recent papers have examined the various compounds
found in blaeberry. Zhang et. al. (2004) compared different HPLC methods
(HPLC-DAD and HPLC-MS) in an attempt to develop better methods for
distinguishing between blaeberry anthocyanins. Their conclusion is that HPLC
analysis before hydrolysis is very useful for controlling product quality in
identification and consistency of raw materials, but hydrolysis is better for
quantifying the individual anthocyanidins (Zhang et. al. 691). Du et. al. (2004)
also isolated two anthocyanin sambubiosides using another technique, high
speed counter-current chromatography (HSCCC), which has the benefit over
18
HPLC of high sample loading capacity, no adsorption of the materials to solidphase column material, and complete sample recovery, making it an effective
method for producing pure anthocyanins on a laboratory scale (Du et. al. 2004,
63). Witzell et. al. (2003) examined variation in phenolic compounds for
Vaccinium myrtillus leaves collected from an unfertilized Picea abies forest in
northern Sweden. They established that the concentration of certain compounds
(e.g. p-coumeric acid) varied throughout the growing season (Witzell et. al. 2003,
125). Such temporal variation has obvious ramifications for harvest times,
should these phenolics prove to have properties applicable to industry or
medicine. Zadernowski et.al. (2005) classified phenolics from Vaccinium
myrtillus picked in northeastern Poland. They identified 17 phenolic acids in
Vaccinium myrtillus berries, as compared to five or less in cultivated high-bush
varieties (Zadernowski et. al. 2005, 2120). This has implications because
phenolics are known to be antioxidant, and the quantity of phenolics present in
berries is affected by a number of factors including maturity of the fruit, storage
conditions, and processing methods, amongst others (ibid., 2118). There is
clearly scope for more research on this subject using blaeberries from Scotland.
19
3.1.2 Antioxidant Profiles and Recent Research
Free radicals and reactive oxygen species (ROS) are oxidants that arise in
the human body from various sources including mitochondrial respiration, contact
with environmental toxins, drug metabolism, and cell necrosis. They are
cytotoxic and have been shown to create oxidative stress and inflammation in the
tissues when they accumulate (Donaldson et al. 2003; Brown et. al. 2004; Emerit
et. al. 2001; Stringer and Kobzik 1998). In addition to leading to the natural
process of aging, free radicals/ROS can be implicated in a number of
pathological processes including atherosclerosis, nerve damage, Parkinson’s
disease, cancer, and rheumatoid arthritis. Therefore, encouraging the presence
of natural anti-oxidants, like anthocyanins, in the body to help mop up excess
ROS may help to lower risk factors for these diseases.
Over the last 10 years, research has focused on identifying and
quantifying the antioxidant capacity of anthocyanins. This study found at least
ten scholarly articles on anthocyanin anti-oxidant capacity from extracts of
Vaccinium myrtillus berries in that period. Wang et. al. (1997) used an oxygen
radical absorbing capacity (ORAC)4 assay to determine the antioxidant capacity
of 14 available anthocyanin extracts when compared to anti-oxidant standard and
vitamin E analogue, Trolox. They were able to demonstrate that the five
anthocyanidins commonly present in fruits (dephinidin, cyanidin, malvidin,
peonidin, and pelargonidin) have varying rates of ORAC capacity based on the
hydroxylation of their B-rings, whether they were glycosylated, and the type of
glycosylation (e.g. glucose v. rhamnose) for each molecule. The cyanidin-3glucoside had the highest anti-oxidant capacity, and pelargonin had the lowest of
all the anthocyanins tested (See Figure Eight).
4
Oxygen radical absorbing capacity (ORAC) is a method for determining the
ROS inhibition capacity for a number of compounds varying from animal tissue to
fruits and vegetables.
20
Figure Eight: Order Ranking of Anthocyanins by ORAC Capacity
Compound
Common Name
Present in Vaccinium
Percent of Total
myrtillus
Anthocyanins in
Blaeberry
1
cyanidin-3-glucoside
kuromanin
2
cyanidin-3-rhamnoglucoside
keracyanin
3
cyanidin
4
cyanidin-3-galactoside
5
malvidin
6
delphinidin
7
peonidin-3-glucoside
8
cyanidin-3,5-diglucoside
9
peonidin
10
pelargonidin-3-glucoside
callistephin
11
malvidin-3,5-diglucoside
malvin
12
pelargonidin
13
malvidin-3-glucoside
oenin
14
pelargonidin-3,5,diglucoside
pelargonin
ideain
cyanidin-3-glucoside
8.1%
cyanidin-3-galactoside
8.9%
peonidin-3-glucoside
2.7%
malvidin-3-glucoside
8.2%
cyanin
After Wang et. al. (1997), 306; Dugo et.al. 2001, 3990.; Kahkonen et. al. 2003, 1409.
Similarly, Kahkonen et. al. (2003) and Nakajima et. al. (2004) profiled
anthocyanins from Vaccinium myrtillus and other berries and measured their
antioxidant properties. Nakajima et. al. (2004) measured anti-oxidant activity by
DPPH5 radical-scavenging activity. They found that blaeberries have more
dephinidin and cyanidin glycosides than commercially produced high-bush
blueberries (Nakajima et. al. 2004, 244). Kahkonen et. al. (2003) used fresh,
purchased fruits from Finland. They note that there are many variables that can
affect anthocyanin content of berries including varietal and regional diversity,
growing conditions, humidity, temperature, fertilizer use, and stress factors like
disease (Kahkonen et. al. 2003, 1408). They note in their results that the total
amount of anthocyanins present in blaeberries was similar to that found in other
studies, but that amounts of malvidin-3-glucosides varied (Kahkonen et. al. 2003,
1409). This makes it all the more important that berries from different regions
5
2,2-Diphenyl-1-picrihydrazyl EPR spectroscopy is another method for effectively
determining the ROS-scavenging capacity of a compound.
21
and environments in Scotland be tested for variation in anthocyanin content and
quantity, and the results compared with other studies.
Likewise, a number of other studies have examined antioxidant and antilipid peroxidation by the anthocyanins and phenolics in Vaccinium myrtillus
berries. Faria et. al. (2005) used field-picked blaeberries (origin unspecified) and
free-radical scavenging ability was measured using DPPH and the ferricreducing/antioxidant power (FRAP)6 assay. Significantly, in their study
chemically modified extracts were found to have higher radical scavenging and
anti lipid-peroxidation capacities than pure blaeberry extracts. These results
have potential for maintaining stability in pigments used in the food industry, but
their ramifications for human consumption or health are unknown. Prior et. al.
(1998) and Ehala et. al. (2005) examined antoxidant capacity by phenolic content
(and anthocyanin content in the case of Prior et. al.) in Vaccinium myrtillus
berries from Germany and Estonia. Prior et. al. sampled various Vaccinium ssp.
berries from America and Maine, but ordered their blaeberries from Germany.
They found that the blaeberries and their close relatives the low-bush blueberries
from Canada had the highest antioxidant capacity, but they also concluded that
there was considerable variability amongst all of the Vaccinium spp. samples
(Prior et. al. 1998, 2690) underscoring the necessity to sample regionally in
Scotland and compare these results to other studies. Ehala et. al. (2005), using
fresh berries purchased in Talinn, Estonia, showed that blaeberries were found to
have the highest total phenolic content and antioxidant capacity of all of the
berries they tested. However, all of the studies agreed that much more work
needs to done in order to understand the variability in berry anthocyanins and
phenolics, as this will affect their value as antioxidants in both the food and
phytochemical markets.
6
The ferric reducing antioxidant power (FRAP) assays are a group of assays for
testing the ferric reducing capacity, therefore antioxidant ability, of a number of
biological fluids.
22
3.2 Biomedical Research: Current Trends and New Horizons
Although the earliest medical studies on bilberry extracts were concerned
with whether they affected eyesight and ocular vasculature, more recent work
has focused on possible applications for the antioxidant properties of Vaccinium
myrtillus berries. Medical and pharmacological research seems to fall into a
number of categories amongst which are pharmacokinetic studies, studies on the
cytoprotective qualities of anthocyanins, and studies on the antidiabetic
properties of anthocyanins. Some of the earliest studies focused on
pharmacokinetics, because bio-availability is crucial for blaeberry antioxidants to
be useful in human health and medicine. Lietti and Forni (1976, 834) found that
blaeberry anthocyanins administered to rats moved rapidly from the blood to the
tissues, particularly the kidneys and skin, where they had a long-acting activity on
the capillaries that lasted beyond when the chemicals were no longer detectable
in the blood. However, low bio-availability in rats has been reported due to
hepatic extraction (Lietti and Forni 1976, 835; Morazzoni and Bombardelli 1996,
17), so the mechanisms for this are not understood. More recently, research has
shown that anthocyanins are well absorbed in the stomachs and small intestines
of rats, (Talavera et. al. 2003, 4181; Talavera et. al. 2004). They also found that
the delphinidin glycosides, which make up approximately 40% of the total
blaeberry anthocyanin content, were better absorbed into the animal tissues than
other anthocyanins (Talevera et. al. 2003, 4181). The bio-availability of
anthocyanins in humans is also unknown (Guohua et. al. 2001; He et. al. 2005).
Guohua et. al. (2001) gave elderberry anthocyanins to a small group of elderly
women and were able to measure the unchanged anthocyanins in blood and
urine by HPLC, with a peak at 71 minutes after ingestion. They found that most
of the anthocyanins had passed out of the body after about 4 hours (Guohua et.
al. 2001, 924). More recent studies of adult males (Kay et. al. 2004) have
concluded that anthocyanins are metabolised into other molecules in the human
23
body, which may contribute to or detract from their bioactivity. Research on
anthocyanin metabolism in the human body is not extensive (Kay 2006;
Morazzoni and Bombardelli 1996, 14), sample groups have been very small, and
methodological problems in previous studies have been raised (Kay 2006, 141),
so while it is still thought that anthocyanins have health benefits, more research
on metabolism and pharmacokinetics needs to be done before the health
benefits of anthocyanin ingestion can be confirmed.
Whilst anthocyanin bio-availability is, in general, unknown, their bioactivity is the subject of much current research. Bilberry extracts have been
shown to be protective against oxidative stress in rat hepatocytes (Valentova et.
al. 2007), in aged rats with cognitive deficits related to oxidative stress (Kolosova
et. al. 2005), and in human subjects fed a high-fat diet (Kay and Holub 2002).
One recent paper suggested that the antioxidant activity of blaeberry extract
could be the reason for the improvement of fibromyalgia symptoms in sufferers
participating in a small double-blind, placebo-controlled trial (Logan and Wong
2001, 453). Blaeberry extracts have also been shown to have anti-inflammatory
(Lietti et. al. 1976; Morazzoni and Bombardelli 1996) and wound-healing
activities (Morazzoni and Bombardelli 1996), vasoprotective activity (Boulakia et.
al. 2000; Morazzoni and Bombardelli 1996; Lietti et. al. 1976), Anti-ulcer activity
(Morazzoni and Bombardelli 1996; Chatterjee et. al. 2004), insulin-regulatory
ability (Jayaprakasam et. al. 2004), and lipid-lowering activity (Morazzoni and
Bombardelli 1996; Cignarella et. al. 1996). Ironically, the traditional use of
blaeberry to improve dark-adaptation and night vision has never been clinically
proven. Muth et. al. (2000) performed a double-blind, placebo-controlled trial of
blaeberry anthocyanins on the improvement of night vision in young male
subjects with no significant effect. Canter and Ernst (2004) also reviewed a
number of placebo-controlled trials on night vision improvement in healthy
subjects, and found that there was no evidence to support this hypothesis. On
the other hand, other studies (as cited in Morazzoni and Bombardelli 1996; Mills
and Bone 2000) have shown improvements in diabetic retinopathy and retinal
damage in those with compromised eyesight. All of these applications have
24
potential for development as more research is completed on anthocyanins and
the other phytochemicals present in Vaccinium myrtillus.
One particular field of study remains the ‘holy grail’ of medical research,
and that is the effect of phytochemicals, here anthocyanins, on cancer cell
proliferation. Two recent papers on the general subject of anthocyanins and
cancer prevention (Cooke et. al. 2005; Kang et. al. 2003) suggest that
anthocyanins are able to inhibit the growth of cancer cells in vivo, although
Cooke et. al. (2005, 1938) caution that pharmacological potency differences
between different anthocyanins should be established, and tissues in which their
protective ability may be applied should be explored. Olsson et. al. (2004)
examined the inhibition of breast cancer cells by anthocyanins from various fruits,
including the commercial high-bush blueberry, and found that blueberries had
among the highest inhibition effects of breast cancer cells HT-29, although mixed
rates of proliferation were reported. Wisely, they also suggest that the effects on
proliferation may be influenced by the synergistic activity of more than one
compound found in an extract (Olsson et. al. 2003, 7269), an observance that is
not often made in pharmacological studies on plant chemicals. Similarly,
experiments have been performed on a variety of cancer cells to establish
whether Vaccinium myrtillus extracts and other blueberry extracts can induce
apoptosis in these cells. Katsube et. al. (2003) found that delphinidin and its
glycoside extracted from Vaccinium myrtillus inhibited the growth of HCT116
colon cancer cells in vitro by apoptosis and that dephinidin and malvidin and their
glycosides inhibited growth of HL60 colon cancer cells by apoptosis. Yi et. al.
(2005) found that anthocyanin extracts from American commercially grown
blueberries showed the highest anti-proliferation activity in HT-29 and Caco-2
colon cancer cells when compared to other fractions like phenolic acids,
flavonols, and tannins. This is significant because wild blueberries have been
shown to have significantly higher amounts of anthocyanins than high-bush
blueberries (Moyer et. al.2002; Beccaro et. al. 2006), and therefore may prove to
have more effective anti-proliferation activities on the these cells lines. Finally,
Zhao et. al. (2004) found that anthocyanin rich extracts (AREs) from several
25
berries including Vaccinium myrtillus inhibited growth of colon cancer cells HT29, but did not inhibit non-tumorigenic cells. This is significant in an environment
like the colon, where high cell turnover plays a role in tissue health.
As a final point, two studies from 2005 focused specifically on how
anthocyanidins and anthocyanins work on a molecular level in COX-27
production and colon cancer (Hou et al. 2005; Lala 2005). Hou et al. (2005)
used anthocyanidins on LPS-induced RAW264 murine macrophages to observe
COX-2 expression in vitro. Their study showed that delphinidin and cyanidin,
had dose-dependent COX-2 inhibition at mRNA and protein levels. This
experiment also showed that this was due to blocking the MAPK-mediated
pathway including activation of inflammation mediator proteins NF-κB and AP-1.
Geeta Lala (2005) studied the bio-availability and chemoprotective activity of
anthocyanin rich extracts (AREs) on an Azoxymethane-induced rat colon cancer
model. She found that, in confirmation of in vitro experiments on colon cancer
cell line HT-29, COX-2 mRNA was significantly down-regulated in rats fed AREs.
The importance of Lala’s research is two-fold: it is the first known study in which
inhibition of colon cancer cell proliferation by bilberry AREs has been shown in
vivo (Lala 2005, 56), and by using AREs, she demonstrates that the ingested
anthocyanins are, in part, responsible for the chemoprotective effect. It remains
unknown whether the anthocyanin aglycones used by Hou et al. (2005) are bioavailable in animals after berry consumption, in the lab they must be produced
chemically. As chemical assays and equipment become more complex and the
molecular mechanisms of disease are better understood, it seems likely that
there will be more novel applications for bilberry extracts within the occidental
and herbal medical establishments.
7
Cyclooxygenase-2 is an enzyme mediator of prostaglandin synthesis in the
body. Prostaglandins perform a number of important duties in the body including
regulating inflammation and protection and repair of tissues.
26
Chapter Four: Vaccinium myrtillus as an ‘Industry’
4.1 The Role of Vaccinium myrtillus in the Wild Food Industry
The Non-Timber Forestry Products (NTFP) industry is a developing
industry in many countries, including the Scandinavian countries and Eastern
Europe, although many would argue that people have been using the forests to
supplement their needs for millennia. This categorization is a catch-all label for a
large number of fungi, wild plants, and plant-related products that are the subject
of growing interest from academics, rural developers, and industries across the
world (Emery et. al. 2006, 1). NTFPs are also occasionally referred to as special
forest products (Molina et. al. 1997) or non-wood forest products (NWFPs)
(Olmos 1999; Saastamoinen 1999), and include the berries, flowers, stems,
twigs, seeds, nuts, or bark (and products made from these) of many plants that
grow in a forested or wild environment. Mushrooms and lichens are also included
in this category. Interest in NTFPs has generated many recent studies by a
variety of international, national, and local groups including the United Nations
and other international development agencies like the NTFP Exchange
Programme for South and Southeast Asia, the United States Department of
Agriculture Forestry Service, the Royal Botanic Gardens at Kew and Edinburgh,
the Forestry Commission, Reforesting Scotland, and other quangos and interest
groups. The current surge in research about NTFPs in the UK is being driven by
government and Forestry Commission interest, even though the NTFPs,
themselves, may have no direct link to forestry (Douglas Hardie, Highland
Natural Products, pers. comm. 24/4/07). For instance, new work funded by
Forestry Commission Scotland and Scottish Enterprise and carried out by
Reforesting Scotland, is currently assessing the possibility of creating a trade
body for the support of NTFP businesses in Scotland8. There is currently too
8
Links to this and a number of other research projects on NTFPs in Scotland are
available from http://www.forestharvest.org.uk/projects.htm.
27
much research taking place in this subject area for it to be covered in detail in
this work, but it is important to discuss recent NTFP research on blaeberries here
and abroad.
In many European countries, NTFPs are free to the general public
provided people harvest and process them without damaging the forest
ecosystem (Saastamoinen 1999), but increasing use of NTFPs on an industrial
scale has been cause for conflict and discussion in more than one European
country. Sustainability, along with access rights and ownership, are among the
topics that NTFP interest groups are now debating. Recently, a number of
studies have been published about NTFPs across the globe, and many of these
include reference to blaeberry collection. A study for the Russian Far East
(Khaborovsk Region) funded by the Canadian International Development Agency
found blaeberries to be one of the most popular berries harvested in the region,
and that numerous processing companies exist for turning them into jam that is
mainly used for Russian consumption (Sprieszl 2001). Similarly, the USDA
Forestry Service (1999) commissioned a study to investigate the potential for
marketable NTFP products from Alaskan forests. Although this was largely a
paper examining NTFP policies in America and Canada, 70% of the businesses
interviewed for the study were seeking new sources of blaeberry leaf and berry at
the time. Demand quotes from these businesses for Vaccinium myrtillus were
approximated at 10 to 20 tonnes per annum for dried leaf, and 5 to 10 tonnes per
annum dried berry with quoted prices of $3/kg for berry and $3.20/kg for leaf. It
was claimed that the demand for dried leaf was particularly high, which has the
potential of adding value to the crop if harvesting and processing methods for
leaf can be established in Scotland. In Project Blaeberry, Fiona Sinclair (2000)
reports on European and Scandinavian markets, where Sweden and Finland
remain the top producers of wild berries for the export market. Sweden averages
a harvest of 485 million kg of wild berries (including blaeberries, lingonberries,
and cloudberries) per annum, and Finland averages 450 million kg, but can
produce 730 million kg in a good year (Sinclair 2000, 7). It appears that, like in
28
Russia, most of the berries are harvested and individually quick frozen (IQF) for
the jam industry, but that berry concentrates for drinks are also produced (ibid.).
Here in the UK, Vaccinium myrtillus berries as NTFPs are also the subject
of much research. Sanderson and Prendergrast (2004) were commissioned by
Kew Gardens and The Countryside Agency to look at the harvest of ‘wild foods’
in England and Scotland. They report the annual harvesting of ‘whinberries’ in
Shropshire, sales of which are known to top £10,000 per annum (Sanderson and
Prendergrast, 56). They quote a market price of £5.50/kg paid for blaeberries in
Shropshire in the year 2000. They also make some important observances
about the costs and difficulties of harvesting wild foods in the UK, including
problems with an infrastructure, which is largely non-existent in most places, and
the seasonality of crops, which affects collectors and supply to industry.
In
Scotland, blaeberries as wild foods are also very much on the research agenda.
Over a decade ago, Agros Associates produced a report for Highlands and
Islands Enterprise outlining the possibilities for harvesting a number of wild plant
products in Scotland. They project the total market outlet for Vaccinium myrtillus
to be 50 tonnes annually for berries and 6 tonnes annually for leaf, for Europe
alone. Notably, since this report was written, the healthcare applications for
bilberry anthocyanins have grown exponentially based on new research into antioxidants, and it would be interesting to recalculate these figures for the current
market.
More recently, Fiona Sinclair’s (2000) Project Blaeberry, funded by the
Millennium Forest for Scotland Trust, had the purpose of studying the
possibilities of inter-cropping blaeberries in a forest environment, where they fruit
best. It remains a very important monograph of Vaccinium spp. research until
the year 2000, although there is little information within on the applications of
such research to the actualities of blaeberry management, harvesting, and
processing in Scotland. The Scottish Executive also funded a 2001 study on the
sustainable development of Scottish plants for industry. Whilst berries in general
are discussed as a possible development area with regards to the food and
drinks industry, the report contains no new information on Vaccinium myrtillus
29
development for any industry, especially herbal medicine. Finally, the NonTimber Forest Products Scotland group and their associates continue to sponsor
research9 and seminars on developments in this growing area of production, and
future work on a ‘blaeberry industry’ will certainly require regular communication
with them as projects develop.
9
For recent NTFP research projects see
http://www.forestharvest.org.uk/projects.htm.
30
4.2 Vaccinium myrtillus Crop Trials
High-bush blueberries have been cultivated in Europe since the
early part of the 20th century, but even today there is very little information on
cultivation of Vaccinium myrtillus from anywhere in the world. In his overview of
Vaccinium ssp. cultivation in Europe, Pliszka (1997) noted that work on blaeberry
cultivation has taken place in Slovenia, but unfortunately no further details about
this research were given. A Ph.D. student at the Swedish University of
Agricultural Sciences, Andreas Akerström, is currently running crop trials on
Vaccinium myrtillus. He is interested in the cultivation of blaeberries because,
amongst other reasons, Polarica AB10 has decided that there is no longer a
future in wild-harvesting berries while the market continues to grow (Andreas
Akerström Profile: http://www.hfon.org/fon/engelska/eng-doktorander.htm). The
author did attempt to contact Mr Akerström about his research, but he has yet to
return any emails, so the outcomes of his study are currently unknown.
The Australian Government recently published a study on blaeberry trials
that they have been running in Tasmania (Australian Government RIRDC 2006).
They had interesting results that may benefit future crop trials here in Scotland.
As there is no Vaccinium myrtillus native to Tasmania, and quarantine rules and
fees are prohibitive for imported plants, they had to germinate seed themselves.
Initially, they had a low germination success rate, even with stratification, so they
instead propagated by cultivating leaf cuttings, buds, and stems, which they
found was not practical for large-scale plantings. Subsequent germinations with
viable seed produced 1000 seedlings for trial at two sites, one in a mild coastal
climate, and one in a cooler area at an altitude of 615m. The soil was acidified
and fertilised, and plants were monitored for 3 years. The plants flowered in the
10
Polarica AB is Europe’s largest supplier of berries and has been in the berry
business for 25 years. They have considerable knowledge and expertise in the
area of purchasing and processing of wild berries.
31
third year, when the plants were 32 months old. There was fruit-set at altitude,
but none set in the milder climate. Additionally, the plants at the higher altitude
grew 50% larger than those in the lower, more temperate field. Additional
cultivation information arose from the author of the RIRDC study travelling to
Poland to research the growth and harvest of blaeberry in the forests there. He
discovered from his Polish colleagues that blaeberry was sparse in forests where
deciduous species dominated, which may be the case in Scotland, as well. In
Polish forests, the best blaeberry stands were found in 50% canopy cover and
blaeberry grows relatively slowly there, gaining only 14% cover after 50 years
(Australian Government RIRDC 2006, 26), information that may prove useful for
the cultivation of blaeberry in Scotland.
Another recent study of Vaccinium myrtillus use in the Ukraine
(Konovalchuk and Konovalchuk 2006, 56) found that, there, blaeberries grow
best in pinewoods, mixed birch and pinewoods, and mixed oak and pinewoods.
The Ukranian study estimated crop yields of around 80 kg per hectare, but they
report that other studies have found this to be 110 kg per hectare with roughly
half of that being suitable for harvesting11 (ibid.). They also recommend tree
felling in the winter, when blaeberry plants are dormant, and they advise the use
of organic fertilisers, herbicides, and pesticides for improved husbandry. Parlane
et. al. (2005) studied growth of Vaccinium myrtillus in Pinus sylvestris woodland
in Strathspey in Scotland, with relationship to light levels and herbivorous
browsing. They found that blaeberry cover was best in open stands, but rare in
dense plantings, and that browsing reduced cover suggesting the need to control
herbivores to encourage blaeberry growth (Parlane et. al. 2005, 276). They
determined that blaeberry growth was optimal at an irradiance of 0.35, and they
recommend controlling tree height to encourage establishment (ibid.).
11
There are other studies on crop estimation for Vaccinium myrtillus berries.
Ihalainen et. al. (2002) created mathematical models of yield based on current
forestry methods, site, and stand characteristics in Finland which might prove
useful in creating similar predictions for Scottish forests.
32
The author could find no literature to suggest ongoing trials or cultivation
of Vaccinium myrtillus in North America. However, the native wild blueberry
Vaccinium angustifolium is grown extensively in Maine and along the east coast
of Canada into Nova Scotia, providing up to 50% of the total blueberry crop for
North America (Yarborough 2002, 409). Yarborough (2002, 410) surveyed fields
of wild North American blueberries (Vaccinium angustifolium) that had been
cultivated from wild stands, with the intention of creating a better model for
estimating crop yield. He found considerable clonal variation in all of his sampled
locations. He also discovered that by current methods, all estimates of berry
yield were considerably higher than the average yields by nearly 40%. He
attributes this to issues with estimating field cover. Fiona Sinclair (2000) visited
wild blueberry growers in Canada for Project Blaeberry. She found that the
farmers maintained their wild blueberry fields by mowing or burning in a two-year
cycle. This apparently has the knock-on effect of producing more fruit. She also
learned that wild blueberry fields often took up to 10 years to establish by
rhizomatous spread, and that they were generally burnt every 6 years to keep
down weed growth. In 1997, Yarborough noted that 52,800 hectares of wild
blueberries were cultivated in North America. He, like Sinclair, found that adding
bee hives to the fields, reducing weed competition, and developing appropriate
fertilising methods greatly increased wild blueberry yield (Yarborough 1997, 34).
Yet, because of clonal differences within crops, wide variations in fruiting do exist
in wild species, and continuing research into blossoming, fruit set, and berry size
amongst clones is necessary to improving the yield in these crops (Smagula
1997, 113). Certainly, continued study of the cultivation techniques of these
blaeberry cousins growing in America and Canada may provide some insight into
better ways to manage blaeberry cultivation in Scotland.
In 2002 Highland Council and the local enterprise companies in
association with Colin Stirling of HortiCS began a project to determine if certain
berry crops could be established and grown intensively (Stirling 2006, 2) at a
number of sites in the highlands. A three-year crop trial was established at 14
sites. These included Skye, Arisaig, Halkirk, Bonar Bridge, Borgie Forest, and
33
Helmsdale (Sarah Allen, pers. comm. 24/04/07) for cultivation of Vaccinium
myrtillus. Site holders were expected to prepare the site for cultivation and the
plants came from four sources: the Clashnadarroch Forest (Aberdeenshire),
North Sutherland, Alba Trees Ltd. (Borders), and from Finland where they had
been produced by micropropagation. Some blaeberries were planted in an
exposed site, and suffered badly from wind damage throughout the trial. As this
is a forest species, it would make more sense to cultivate them in a forest
environment, and this was noted in the report (Stirling 2006, 4). The report
concluded that blaeberries would take at least 5 years to establish as a crop, but
this author feels that if a comparison is made to Vaccinium angustifolium crops in
Canada, this could, in actuality take as long as 10 years (Sinclair 2000).
Unfortunately, the official report on these trials was very brief and contained no
actual data. Any extended blaeberry project would need to see further
investigation into the trial data. The project coordinator did compile some short
leaflets (See Appendix Four) with cultivation instructions for the Highland Berry
Growers Group, but only extended trials will determine the optimum environment
necessary for effective blaeberry cultivation. The author does not know what has
happened to the sites since the trial was finished in 2005, the contact addresses
the author was given are no longer functioning, and the author has been unable
to reach any of the people involved with the trials. A brief chat via email with Ella
Drinks managing director Anne Thompson revealed that the project had gone
quiet, simply because there was no one currently willing to coordinate the
ongoing research (Pers. comm. 11/04/07). There is plenty of scope for further
investigation, here, however. Extension of this research will allow more time to
find and communicate with the project participants and, perhaps, examine the
data from the trials and visit the sites to see if any valuable information might be
gained from the crop trial study.
34
4.3 Harvesting and Processing
The blaeberry industry continues to grow worldwide because of demand
for the berries for food, beverages, condiments, and medicines. Yet there are
many logistical issues that dominate potential crop development, including
harvesting and processing methods, extract production, and shipping and
marketing, to name a few. Harvesting is an almost universal problem for
industrial scale production of Vaccinium myrtillus berries. It is the greatest
expense in producing the wild blueberry crop in America (Yarborough 2002,
329), where most of it is still collected by hand (Sinclair 2000, 12) using the hand
rake or a similar tool (See Figure Nine).
Figure Nine: Collecting Blaeberries in Clashwood, Ross and Cromarty
Taken by the author July/August 2006
It is the author’s supposition that this is one reason that berry purchasers and
processors like Polarica AB are no longer interested in wild-harvesting berries;
the cost of buying through agents that have collected berries from individuals at
collection stations (Leven and Segerstedt 2004) has most likely become
prohibitive in a rapidly expanding market. If Scotland is to finally develop
35
Vaccinium myrtillus as an agricultural crop then considerable resources will have
to be put into developing a mechanical harvester that does not crush the fruit,
and can work in a woodland environment. Recent development of a harvester
uniquely designed for the harvest of Bog Myrtle (Myrica gale) in a collaboration
between Cranfield University, Highland Natural Products, and LogLogic has
shown that cooperation between industry and academia can produce new
technology specifically designed for unique harvesting situations. Yarborough
(2002) discusses the progress towards developing a wild blueberry harvester for
North American crops. He reports that the most commonly used mechanical
harvester for wild blueberries is the Bragg harvester (See Figure Ten), but that in
trials this only recovered between 59% and 69% of berries as compared to hand
harvest (Yarborough 2002, 330).
Figure Ten: A Bragg Blueberry Harvester on a Tractor
http://www.wildblueberries.net/harv.html
However, during recent trials, use of a prototype harvester by Nimco with canoeshaped teeth that strip the berries without damaging the plant, eliminated the
problem of crushing of berries that happens with reel-type harvesters. The
picking head was also monitored by a set of sensors that allowed it to move over
hummocks and obstacles, preventing damage to the head, which can often
happen on rocky terrain. This harvester performed as efficiently as handharvesting in most cases, but it was slower than other harvesters because it was
a prototype and not suitable to be monitored for harvesting efficiency
36
(Yarborough 2002, 331). As with the other research areas discussed in this
work, it seems clear that under the right circumstances there is plenty of scope
for successful development of new technology for large-scale harvesting of
blaeberry, even if it is being cropped in a wooded environment.
Post-harvest processing of blaeberries is another area where there has
been significant research. Vaccinium myrtillus berries, and their constituents, are
fragile and must be properly preserved by freezing, drying, or cooking within 48
hours of picking (Leven and Segerstedt 2004). Polarica AB, one of the world’s
largest processors of blaeberries, uses freezing tunnels to freeze the berries that
can then be packed into 500 kg boxes and stored (ibid.) for future sorting or
shipment. This practice is only limited by freezer space and by how rapidly
berries arrive at the processing plant during the short picking season. Berries
can also be individually quick frozen (IQF), a technology also used for seafood
and other foods, spray-dried, heat-dried or cooked, but most of these will require
the processor to provide space for expensive equipment, and a food-standard
production environment. Research in this area is very much focused on
treatments that will improve the quality and ripeness of the fruit, reduce waste
during processing, reduce infection by biological contaminants, and preserve the
quality and efficacy of the juice or extract. Please see the bibliography
(Appendix One) for a more resources on this subject.
Preparing anthocyanin extracts from blaeberry is a complicated and
expensive process that requires several steps and expensive laboratory
equipment:
The first step in the manufacturing process is the biological elimination of
the sugars by Saccaromyces spp.; the medium recovered from the
fermentors is clarified by on-line centrifugation and concentrated under
vacuum at low temperature. The purification of this crude extract entails
repeated treatment with boiling ethanol. Upon cooling of the ethanol
solution, the insoluable anthocyanins precipitate and are recovered by
centrifugation (the free anthocyanidins, organic acids, and other impurities
remain in solution (Bruneton 1999, 362).
However, more recent techniques like sub-critical water extraction (SWE) are
providing cheaper and more efficient methods for producing anthocyanin
37
extracts. King et. al. (date unknown)12 working through the Los Alamos National
Laboratory in New Mexico have developed a method for extracting anthocyanins
without ethanol or other solvents by using the SWE technique, which alters the
boiling point of the fluid (water) under pressure, changing its properties and
allowing it to extract polyphenolic molecules from berries (King et. al., 3).
Anthocyanin extraction from elderberries (Sambucus nigra) and chokeberries
(Aronia ssp.) by SWE produced equivalent or better extraction of anthocyanins
than by ethanol (King et. al, 4). They conclude that SWE would be a good
technique for extracting anthocyanins cheaply and efficiently, and it would prove
very interesting to see how well the technique worked for blaeberry
anthocyanins. Another recently developed technique for extraction of plant
polyphenolics and antioxidants is supercritical fluid extraction (SFE) particularly
by supercritical CO2. A brief survey of internet-based businesses selling
blaeberry extracts, seemed to indicate that this was a common way to produce
pharmaceutical-grade blaeberry anthocyanins, but a similar survey of scholarly
articles on the Ingenta Sciencedirect website did not being up any relevant
articles (search terms: supercritical fluid extraction + bilberry; supercritical fluid
extraction + anthocyanin). SFE-CO2 has until very recently been a very
expensive way of producing extracts (Douglas Hardie, pers. comm. 25/04/07).
More research into these techniques and other current research will be
necessary as a blaeberry project develops, but it is not possible within the current
scope of study to develop this topic any further.
12
This paper was found on the website of the Los Alamos National Laboratory
website, http://scrub.lanl.gov/index2.htm, but its publication date is unknown. The
website was first published in 2004.
38
Chapter Five: Discussion and Conclusion
5.1 Discussion
When the preliminary research plan for this study was undertaken in May
of 2006, it was to encompass a thorough literature review of the subject of
Vaccinium myrtillus cultivation for use in food, drinks, and healthcare, and an
extended monograph of the plant including current research from the last 10
years on its industrial use and cultivation in Europe and Scotland. A year later, it
has become clear that such a project was naïvely well intentioned, but
completely impossible within the time frame and scope established for this
dissertation. The literature alone has taken many hundreds of hours to locate
and sift through, much less to read and analyse. However, the benefits of taking
on such an unwieldy project have also become apparent. Blaeberry research is
on the rise, and so much groundwork has already been done, that it seems
possible that work on formalizing blaeberry production in Scotland could really
begin in earnest if the proper infrastructure were put into place. After much
consideration of the literature presented here, it is possible to create a catalogue
of subjects that should be addressed in order to make blaeberry viable as a
commercial crop.
1. Botany and Ecology
a. Continuing research into several aspects of Vaccinium myrtillus
ecology will be necessary to effective crop production. This will
include pollination research for Scottish plants in their natural
habitat and in test ‘fields’, even if these are wooded. The research
should examine which insect species pollinate blaeberry in Scottish
woodlands, its levels of self-pollination, and fruiting success rates
between kinds of pollination. Studies of the environmental impact
blaeberry pollination and fruiting would also prove useful. These
could include the effects of temperature, wind, light, soil moisture
and acidity, fertilisers and pesticides, weed growth, and even
genetics on successful reproduction.
39
b. Continuing research into mycorrhizal associations between Scottish
blaeberries and other forest species may help to elucidate the
relationship between these species and its effect on successful
growth and nutrient uptake. If Vaccinium myrtillus has optimal
growth in Pinus sylvestris woodlands, is this related to their shared
mycorrhizal association (MacKay 2004)? Would mycorrhizal
associations be necessary to improve nutrient uptake in cultivated
blaeberry? How would crop maintenance (e.g. burning) affect the
mycorrhiza? How do they regenerate? Perhaps a mycorrhizal
specialist at RBGE or an agricultural college would be interested in
pursuing this line of study.
c. Continuing research into reproduction, both sexual and clonal,
would be very important to establishing blaeberry as a fruiting crop
in Scotland. Recent reports (e.g. Stirling 2006) did not include
coordinated data to show which clones or seedlings were
established at which trial site, and of these, which clones or
seedlings did best in growth and flower. On the scant evidence
available, clonal reproduction seems to be the best candidate for
large-scale production of plants. Growers would need to know if
clones are more likely than seedlings to successfully establish in
large scale planting. It would be helpful to know, as well, if
rhizomatous propagation might reduce cost of planting on a large
scale. Some of the issues surrounding using a single clone for crop
establishment have been presented in Appendix Two, and
masting behaviour may also be a very big problem with regular
annual harvests. Using multiple clones may be necessary. If
companies like Alba Trees have stocks of plants, they may have
developed efficient ways of germinating or propagating them.
Investigations into which companies have developed methods for
propagating Vaccinium myrtillus will be necessary, as will
determination of potential problems with genetic inbreeding and
masting behaviours. Finding a nursery willing to take on this area
of the project would be ideal.
2. Chemistry
a. Most of the chemistry research that has taken place on Vaccinium
myrtillus extracts has taken place outside of Scotland, in Japan,
America, Italy, Sweden, and Finland. Although chemical
experiments have used a variety of samples from different sources,
very little research has been done chemically on Vaccinium
myrtillus in Scotland. The Berry Scotland website lists recent
research into anthocyanin anti-oxidants in Scotland
(http://www.berryscotland.com/Recent%20research.htm), and this
might offer a route to finding university departments interested in
analysing Scottish blaeberries for their constituents. Ideally, plants
40
and berries should be sampled at different times in the season and
tested for their various constituents. One could even compare
samples from different regions of Scotland with samples from other
countries to see if constituents are present in similar quantities.
b. Anti-oxidant research would need to take place on the Scottish
blaeberry extracts, because, as the above literature review shows
(Chapter 3.1.3), there is considerable spacial and temporal
variation in the antioxidant content of Vaccinium myrtillus berries.
The ORAC and FRAP assays are not expensive and most
laboratories will be able to undertake these assays.
c. It would also be worthwhile to create links with bio-medical
researchers and perform in vitro and in vivo experiments on
possible health applications for Scottish blaeberry extracts. This
kind of research is taking place abroad, particularly in Finland,
Japan, and America, with increasing frequency. Without this kind
of study, Scotland runs the risk of falling behind in developing new
applications for blaeberry extracts and the techniques necessary to
produce them. Napier University School of Life Sciences has a
number of professors and postgraduate students whose expertise
lies in antioxidant research and molecular mechanisms of
pathology induced by oxidative stress. There is scope here for
linking with Napier or other universities to undertake some of this
research for the benefit of a blaeberry development project.
3. Non-Timber Forest Products
a. Full advantage should be taken of the current trend of funding
research into NTFPs. Since Vaccinium myrtillus is likely to grow
and fruit best as a woodland crop, creating links with forestry and
forest owners will be beneficial to running trials, taking samples for
testing, or even wild-harvesting. As many forest owners are now
trying to add value to their woodlands, using timber extraction,
rather than clear-felling, and encouraging blaeberry growth at the
optimum canopy density has the potential to increase the value of
woodlands. Olmos (1999) noted that, at the time of his research,
blaeberries picked in one season from one hectare of Polish forest
were worth nearly the same amount in money as the timber
extracted from one hectare of woodland in a whole year.
Obviously, market values would need to be established for
blaeberry plant products, but this may prove and excellent incentive
for forest owners to work in partnership with a blaeberry crop
project.
b. Legislation and ownership rights for NTFPs need to be examined,
as these are not clear at the moment (see
41
http://www.forestharvest.org.uk/owners.htm). It is likely that a
system for regulating the collection of forest ‘crops’ would need to
be implemented, as has been done in England. Decisions should
also be made about the mapping out of blaeberry resources in
Scotland. Surveys of blaeberry yields over a number of years will
be also be necessary to determine mast cycles, optimal fruiting
environments, and ecological relationships. Sustainable harvesting
practices will also need to be established. Any ongoing blaeberry
project should maintain close links with the NTFP Scotland group
established recently.
4. Blaeberry Crop Development
a. Creating a LINK-type project between academia, agriculture, and
industry would be the ideal way to manage development of
blaeberry as an agricultural crop. Some feasibility studies (Sinclair
2000) and crop trials (Stirling 2006) have already taken place, but
the data for the trials need to be examined more closely. A number
of areas for study have arisen from past crop trials, including the
appropriateness of field sites, propagative techniques, tree cover in
fields, temperatures during the flowering and fruiting season, soil
water levels, light levels, the use of fertilisers and pesticides on
crops, mycorrhizal interactions, and pollination methods, to name a
few. From the analysis of data from recent trials in Scotland, and
data from ongoing crop trials in Sweden and Tasmania, it would
become possible to consider how to move forward with blaeberry
as a crop. A mechanical harvester prototype would need to be
designed, especially if blaeberry were grown as a woodland crop.
Processing infrastructure would also need to be developed. Berries
do not keep more than 48 hours, and during the harvesting season
potentially thousands of tonnes of berries per day would need to be
shipped from field to processing plant. Industry, including food,
drinks, and healthcare might be persuaded to invest in helping with
this research.
b. The author contacted several Scottish businesses via email that
sell or use wild foods in their products, and only one responded.
They were also unwilling to share any information about their use of
blaeberries or potential volume needs for Scottish berries in their
business. Unless industry can be persuaded to participate in
blaeberry research, it will be very difficult to calculate market values
for the crop and weigh those against development costs. It is also
likely to take five to ten years to develop blaeberry as a crop, so
discussions about wild-harvesting potential should be taken in the
meantime. Those involved in previous research, like Fiona Sinclair,
should also be interviewed about their work. The author contacted
Fiona Sinclair by phone, but has yet to be able to organise an
42
interview. Interviews with the Highland Berry Growers Group would
also prove very interesting, as would an interview with Colin Stirling
of HortiCS who coordinated the most recent crop trials. To date,
the author has been unable to contact Mr. Stirling via email or by
phone. Emails to professor Ulla Bäng and Ph.D. student Andreas
Akerström of the Swedish University of Agricultural Sciences also
remain unanswered. Extension of this project will allow time to
make contact with them and possibly find out about their research.
c. Finally, market research will need to be undertaken to identify
potential buyers for Scottish blaeberries. It was originally the
intention of the author to include this subject area in this
dissertation but time and space constraints have prevented it from
happening. Some Scottish food and drink businesses were
approached, but there are likely to be other types of businesses
who might also be interested including herbal tea producers,
healthcare products producers, herbal tincture makers, etc. It
would also be useful to trace supply chains from field to processor
to producer for a number of industries, if permissible. This would
allow a blaeberry project to establish the range of products that
blaeberries could be used for, and which of these would be the
most lucrative.
43
5.2 Conclusion
Despite the overwhelming scale of this project, it has been a most
interesting task to examine the state of research on Vaccinium myrtillus in so
many fields of study. There is scope for development in every area, and the right
individual, in cooperation with business, government, and academia will have
every chance of moving forward with a blaeberry development programme for
Scotland. One thing has become very clear from this study: the project needs a
leader, or coordinator. This should be someone who has the right links, and who
is willing to put in the hard work that will be required to follow up the many
strands of evidence uncovered here. Additionally, much more research needs to
be done, especially on the industrial aspects of harvesting, processing, and
extractions. There will need to be consultations with freezer and processor
suppliers and engineers, experts in developing infrastucture, funding
programmes like the Northern Peripheries Project, and perhaps SEERAD or
other governmental departments and quangos responsible for developing
programmes and protocols in agriculture and the countryside. The author
recommends that surveys of Vaccinium myrtillus cropping and ecology start
immediately, even if mapping and collecting are performed on small scale for a
couple of years. One Scottish business that the author spoke to has expressed
an interest in supporting such a venture this summer. Most importantly, the
momentum building behind this project should be sustained. There is growing
media interest, in the form of national radio and print, on blaeberries as a
medicinal and food plant in Scotland. The author has had one interview about
this work, and has had interest from radio for another possible spot this summer.
This interest should be used to push forward a blaeberry project, rather than
letting it fall silent again, as it has since 2005 crop trials finished. It is a project
full of promise, but the work and investment have only just begun.
44
References
Albert T, Raspe, O., Jaquemart A-L. (2004). Clonal Diversity and Genetic
Structure in Vaccinium myrtillus Populations from Different Habitats.
Belgian Journal of Botany. 137 (2), 155 – 162.
Allen, D.E., and Hatfield, G. (2004). Medicinal Plants in Folk Tradition: An
Ethnobotany of Britain and Ireland. Cambridge: Timber Press.
Australian Government Rural Industries Research and Development
Corporation (2006). Medicinal Herbs – A Preliminary Evaluation of
Astragalus, Bilberry, Feverfew, and Stevia. Tazmania: AGRIRDC,
(05/185).
Beccaro, G., Mellano, M.G., Chiabrando, V., Bounous, G. (2006). Phenolic and
Anthocanin Content and Antioxidant Activity in Fruits of Bilberry
(Vaccinium myrtillus L.) and of Highbush Blueberry (V. corymbosum L.)
Cultivars in North Western Italy. International Society for Horticultural
Science: Proceedings of the VIIIth International Symposium on Vaccinium
Culture. Acta Horticulturae. 715, 553 - 557. Available Online to
subscribers: http://www.ishs.org/acta/index.htm.
Beith, M. (1995). Healing Threads: Traditional Medicines of the Highlands and
Islands. Edinburgh: Birlinn, Limited.
Boulakia, F.C., Valensi, P.E., Boulahdour, H., Lestrade, R., Dufour-Lamartinie, JF., Hort-Legrand, C., Behar, A. (2000). In Vivo Sequential Study of
Skeletal Muscle Capillary Permeability in Diabetic Rats: Effect of
Anthocyanosides. Metabolism. 49 (7), 880 – 885.
Brielmann, H.L. (1999). Phytochemicals: The Chemical Components of Plants.
In, Kaufmann, P.B., Cseke, L.J., Warber, S., Duke, J.A., Brielmann, H.L.,
Natural Products from Plants. London: CRC Press LLC.
Brown, D.M., Donaldson, K., Borm, P.J., Schins, R.P., Dehnhardt, M., Gilmour,
P., Jimenez, L.A., Stone, V. (2004). Calcium and ROS-meditated
Activation of Transcription Factors and TNF-alpha Cytokine Gene
Expression in Macrophages Exposed to Ultrafine Particles. American
Journal of Physiology, Lung Cellualr and Molecular Physiology. 286, L344
– L353.
Bruneton, J. (1999). Pharmacognosy: Phytochemistry Medicinal Plants. 2nd
ed. Paris: Lavoisier Publishing.
45
Canter, P. and Ernst, E. (2004). Anthocyanins of Vaccinium myrtillus
(Bilberry)For Night Vision – A Systematic Review of Placebo-Controlled
Trials. Survey of Opthamology. 49 (1), 38 – 50.
Cao, G., Muccitelli, Sanchez – Moreno, C., Prior, R. (2001). Anthocyanins
Are Absorbed in Glycated Forms In Elderly Women: A Pharmacokinetic
Study. American Journal of Clinical Nutrition. 73, 920 – 926.
Chatterjee, A., Yasmin, T., Bagchi, D., Stohs, S.J. (2004). Inhibition of
Helicobacter pylori in vitro by various berry extracts, with enhanced
susceptibility to clarithromycin. Molecular and Cellular Biochemistry.
265 (1- 2), 19 – 26.
Cignarella, A., Nastasi M., Cavalli E., Puglisi, L. (1996). Novel Lipid-Lowering
Properties of Vaccinium myrtillus L. Leaves, A Traditional Antidiabetic
Treatment, In Several Models of Rat Dyslipidaemia: A Comparison with
Ciprofibrate. Thrombosis Research. 84 (5), 311 – 322.
Cohen – Boulakia, F., Valensi, P.E., Boulahdour, H., Lestrade, R., Doufour –
Lamartinie, J.F., Hort – Legrande, C., Behar, A. (2000). In Vivo Sequential
Study of Skeletal Muscle Capillary Permeability in Diabetic Rats: Effect of
Anthocyanosides. Metabolism. 49 (7): 880 – 5.
Comhairle nan Eileen Siar. (2007). Liostaichean. Available Online:
http://www.cne-siar.gov.uk/gaelic/grd/liostaichean/lusan/lusan_f.htm.
Accessed 28/03/07.
Cooke, D., Steward, W.P., Gescher, A.J., Marczylo, T. (2005). Anthocyans from
Fruits and Vegetables – Does Bright Colour Signal Cancer
Chemoprotective Activity? European Journal of Cancer. 41, 1931 – 1940.
Craig, W. (1879). Milne’s Manual of Materia Medica and Therapeutics.
Edinburgh: E. & S. Livingstone.
Culpeper, J. (1981 (1826)). Culpeper’s Complete Herbal & English Physician.
Barcelona: Harvey Sales.
Dewick, P.M. (2001). Medicinal Natural Products, A Biosynthetic Approach.
Chichester: John Wiley & Sons Ltd.
Dickson C., and Dickson, J. (2000). Plants and People in Ancient Scotland.
Gloucestershire: Tempus Publishing.
46
Dickson, J.H., Oeggl, K., Holden T.G., Handley, L.L., O’Connell, T.C.,
Preston, T. (2000). The Omnivorous Tyrolean Iceman: Colon Contents
(meat, cereals, pollen, moss, and whipworm) and Stable Isotope Analysis.
Philosophical Transactions of the Royal Society London B. 355, 1843 –
1849.
Donaldson, K., Stone, V., Borm, P.J., Jimenez, L.A., Gilmour, P.S., Schins, R.P.,
Knaapen, A.M., Rahman, I., Faux, S.P., Brown, D.M., MacNee, W. (2003).
Oxidative Stress and Calcium Signaling in the Adverse Effects of
Environmental Particles (PM10). Free Radical Biology & Medicine. 34
(11), 1369 – 1382.
Du, Q., Jerz, G., Winterhalter, P. (2004). Isolation of Two Anthocyanin
Sambubiosides from Bilberry (Vaccinium myrtillus) by High-Speed
Counter-Current Chromatography. Journal of Chromatography A.
1045, 59 – 63.
Dugo, P., Mondello, L., Errante, G., Zappia, G., Dugo, G. (2001). Identification
of Anthocyanins in Berries by Narrow-Bore High-Performance Liquid
Chromatography with Electrospray Ionisation Detection. Journal of
Agricultural and Food Chemistry. 49, 3987 – 3992.
Duncan, A. (1829). Supplement to the Edinburgh New Dispensary. Edinburgh:
Bell & Bradfute.
Ehala, S., Vaher, M., Kaljurand, M. (2005). Characterization of Phenolic
Profiles of Northern European Berries by Capillary Electrophosphoresis
And Determination of their Antioxidant Activity. Journal of Agricultural and
Food Chemistry. 53 (16), 6484 – 6490.
Emerit, J., Beaumont, C., Trivin, F. (2001). Iron Metabolism, Free Radicals, and
Oxidative Injury. Biomedical Pharmacotherapy. 55 (6), 333 – 339.
Emery, M., Martin, S., Dyke, A. (2006). Wild Harvests from Scottish Woodlands,
Social, Cultural, and Economic Values of Contemporary Non-Timber
Forest Products. Edinburgh: Forestry Commission. Available Online:
http://www.forestreasearch.gov.uk/fr/INFD-6XCHWF. Accessed: 15/04/07.
Evans, W.C. (2002). Trease and Evans Pharmacology (15th ed ). Edinburgh:
W.B. Saunders Company Limited.
Faria, A., Calhau, C., de Freitas, V., Mateus, N. (2006). Procyanidins as
Antioxidants and Tumor Cell Growth Modulators. Journal of Agricultural
and Food Chemistry. 54 (6) – 2392 – 2397.
47
Faria, A., Oliveira, J., Neves, P., Gameiro, P., Santos – Buelga, C., de Freitas,
V., Mateus, N. (2005). Antioxidant Properties of Prepared Blueberry
(Vaccinium myrtillus) Extracts. Journal of Agricultural and Food Chemistry.
53(17), 6896 – 6902.
Featherstone, A.W. (2002). Blaeberry (Vaccinium myrtillus) Species Profile.
Caledonian Forest Species Profiles. Published by Caledonia Wild!
Available Online: http://www.treesforlife.org.uk/tfl.blaeberry.html.
Accessed: 15/04/07
Forest Harvest. NTFP newsletter. April 2007.
Online: http://www.forestharvest.org.uk/news.htm. Accessed: 10/04/07
Grieve, Maud. (1971). A Modern Herbal: Vol. I, A – H. New York: Dover
Publications, Inc.
Guohua, C., Muccitelli, H.U., Sanchez-Moreno, C., Prior, R. (2001).
Anthocyanins are Absorbed in Glycated Forms in Elderly Women: A
Pharmakokinetic Study. American Journal of Clinical Nutrition. 73, 920 –
926.
He, J., Magnuson, B.A., Giusti, M.M. (2005). Analysis of Anthocyanins in Rat
Intestinal Contents – Impact of Anthocyanin Chemical Structure on Fecal
Excretion. Journal of Agricultural and Food Chemistry. 53, 2859 – 2866.
Honkavaara, J., Siitari, H., Saloranta, V., Vitala, J. (2007). Avian Seed Ingestion
Changes Germination Patterns of Bilberry, Vaccinium myrtillus. Annales
Botanici Fennici. 44, 8 – 17.
Hou, D-X., Yanagita, T., Uto, T., Masuzaki, S., Fujii, M. (2005). Anthocyanidins
Inhibit Cyclooxygenase-2 Expression in LPS-evoked Macrophages:
Structure-Activity Relationship and Molecular Mechanisms Involved.
Biochemical Pharmacology. 70, 417 – 425.
Ihalainen, M., Alho, J., Kohlehmainen, O., Pukkala, T. (2002). Expert Models
for Bilberry and Cowberry Yields in Finnish Forests. Forest Ecology and
Management. 157, 15 – 22.
Jayaprakasam, B., Vareed, S.K., Olsen, L.K., Nair, M.G. (2005). Insulin
Secretion by Bioactive Anthocyanins and Anthocyanidins present in Fruits.
Journal of Agricultural and Food Chemistry. 53 (1), 28 – 31.
Kahkonen, M.P., Heinamaki, J., Ollilainen, V., Heinonin, M. (2003). Berry
Anthocyanins: Isolation, Identification, and Antioxidant Activities. Journal
of the Science of Food and Agriculture. 83, 1403 – 1411.
48
Kang, S-Y., Seeram, N., Nair, M.G., Bourquin, L.D. (2003). Tart Cherry
Anthocyanins Inhibit Tumor Development in APCMin Mice and Reduce
Prolferation of Human Colon Cancer Cells. Cancer Letters. 194, 13 – 19.
Katsube, N., Iwashita, K., Tsushida, T., Yamaki, K., Kobori. M. (2003).
Induction of Apoptosis in Cancer Cells by Bilberry (Vaccinium myrtillus)
and the Anthocyanins. Journal of Agricultural and Food Chemistry. 51 (1),
68 – 75.
Kay, C. (2006). Aspects of Anthocyanin Absorption, Metabolism, and
Pharmacokinetics in Humans. Nutrition Research Reviews. 19, 137 – 146.
Kay, C. and Holub, B.J. (2002). The Effect of Wild Blueberry (Vaccinium
angustifolium) Consumption on Postprandial Serum Antioxidant Status in
Human Subjects. British Journal of Nutrition. 88, 389 – 397.
Kay, C., Mazza, G., Holub, B.J., Wang, J. (2004). Anthocyanin Metabolites in
Human Urine and Serum. British Journal of Nutrition. 91, 933 – 942.
King, J.W., Grabiel, R.D., and Wightman, J.D. (date unknown). Subcritical Water
Extraction of Anthocyanins from Fruit Berry Substrates. Available Online:
scrub.lanl.gov/pdf/king/192_subcritical_water.pdf. Accessed: 27/04/07.
Koenig, W. and Arnold, D. (1961). Facsimile of the First Amsterdam
Pharmacopoaeia, 1636. Nieuwkoop: B. De Graaf.
Kolosova, N.G., Shcheglova, T.V., Sergeeva, S.V., Loskutova, L.V. (2006)
Long-Term Anti-oxidant Supplementation Attenuates Oxidative Stress
Markers and Cognitive Deficits in Senescent-Accelerated OXYS Rats.
Neurobiology of Aging. 27(9), 1289 – 1297.
Konovalchuk V.K. and Konovalchuk V.V. (2006) The Resources of Wild Lowbush
Blueberries (Vaccinium myrtillus L., Vaccinium uglinosum L.) and
Highbush Blueberry culture in Ukraine. In, Lopes da Fonseca, F., Romero
Munoz, F. (eds.), VIII International Symposium on Vaccinium Culture,
ISHS Acta Horticulturae 715. Published Online for the International
Society for Horticultural Science: http://www.actahort.org/.
Korkina, L.G. and Afanas’ev, I.B. (1997). Antioxidant and Chelating Properties of
Flavonoids. In, Sies, H. (ed.), Antioxidants in Disease Mechanisms and
Therapy. In, August, J.T., Murad, F., Anders, M.W., and Coyle, J.T. (eds.),
Advances in Pharmacology, Volume 38. London: Academic Press.
Lala, G. (2005). Effects of Anthocyanin Rich Extracts on Multiple Biomarkers of
Colon Cancer. Thesis (M.Sc.). University of Maryland, College Park.
49
Lang, D.C. (1987). The complete Book of British Berries. London: Threshold
Books.
Leonard, H.C. (1892). Dictionary of Materia Medica and Therapeutics.
London: Bailliere, Tindall, and Cox.
Leven, E. Segerstedt, A. (2004). Polarica’s Wild Berries: An Example of a
Required Storage Capacity Calculation and Where to Locate this
Inventory. Supply Chain Management. 9(3), 213 – 218.
Lietti, A., Cristoni, A., Picci, M. (1976). Studies of Vaccinium myrtillus
Anthocyanosides I: Vasoprotective and Antiinflammatory Activity.
Arzneimittelforschung. 26(5), 829 – 832.
Lietti, A., Forni, G. (1976). Studies on Vaccinium myrtillus anthocyanosides II:
Aspects of Anthocyanin Pharmacokinetics in the Rat.
Arzneimittelforschung. 26(5), 832 – 835.
Logan, A.C. and Wong, C. (2001). Chronic Fatigue Syndrome: Oxidative Stress
and Dietary Modifications. Alternative Medicine Review. 6(5), 450 –
459.
MacBain, Alexander. (1982). An Etymological Dictionary of the Gaelic Language.
Glasgow: Gairm Publications. Available Online:
http://www.ceantar.org/Dicts/MB2/index.html. Accessed: 28/03/07.
MacKay, J.W.B. (2004). Effect of Burning on the Mycorrhizal fungi of Scots Pine
Pinus sylvestris at Boat of Garten, Inverness-shire, Scotland.
Conservation Evidence. 1, 20 – 22. Available Online:
www.conservationevidence.com/Attachments/PDF83.pdf. Accessed:
20/04/07.
Madhavi D.L., Bomser J., Smith M.A.L., Singletary K. (1998). Isolation of
Bioactive Constituents from Vaccinium myrtillus (bilberry) Fruits and Cell
Cultures. Plant Science. 131, 95 – 103.
Mills, S and Bone, K. (2000). Principles and Practice of Phytotherapy, Modern
Herbal Medicine. 1st ed. Edinburgh: Churchill Livingstone.
Moffatt, B. and Fulton, J. (1988). SHARP Practice 2: The Second Report on
Researches into the Medieval Hospital at Soutra, Lothian Region,
Scotland. Edinburgh: Soutra Hospital Archaeoethnopharmacological
Project.
50
Moffatt. B, Thompson, B., Fulton, J. (1989). SHARP Practice 3: The Third
Report on Researches into the Medieval Hospital at Soutra, Lothian
Region, Scotland. Edinburgh: Soutra Hospital
Archaeoethnopharmacological Project.
Molina, R., Vance, N., Weigand, J.F., Pilz, D., Amaranthus, M.P. (1997).
Special Forest Products: Integrating Social, Economic, and Biological
Considerations into Ecosystem Management. Available online:
http://www.fs.fed.us/pnw/pubs/journals/pnw_1997_molina001.pdf.
Morazzoni P., Bombardelli, E. (1996). Vaccinium Myrtillus L.. Fitoterapia. 61 (1),
3 – 29).
Moyer, RA., Hummer, K.E., Finn C.E., Frei, B., Wrolstad, R.E. (2002).
Anthocyanins, Phenolics, and Antioxidant Capacity in Diverse Small
Fruits: Vaccinium, Rubus, and Ribes. Journal of Agricultural and Food
Chemistry. 50, 519 – 525.
Murray, J. (1825). A System of Materia Medica and Pharmacy, Vol. II.
Edinburgh: Adam Black.
Muth, E.R., Laurent, J.M., Jasper, P., 2000. The Effect of Bilberry Nutritional
Supplementation on Night Visual Acuity and Contrast Sensitivity.
Alternative Medicine Review. 5 (2), 164 – 173.
Nakajima, J-I., Tanaka, I., Seo, S., Mamazaki, M., Saito, K. (2004).
LC/PDA/MS Profiling and Radical Scavenging Activity of Anthocyanins in
Various Berries. Journal of Biomedicine and Biotechnology. 2004(5), 241
– 247.
Nuortila C., Tuomi, J., And Laine, K. (2002). Inter-parent Distance Affects
Reproductive Success in Two Clonal Dwarf Shrubs, Vaccinium myrtillus
and Vaccinium vitis-idaea (Ericaceae). Canadian Journal of Botany. 80,
875 – 884.
Nyman, N.A., Kumpulainen, J.T. (2001). Determination of Anthocyanins in
Berries and Red Wine by High-Performance Liquid Chromatography.
Journal of Agricultural and Food Chemistry. 49(9), 4183 – 4187.
Olmos, S. (1999). Non-Wood Forest Products: Utilization and Income
Generation in the Czech Republic, Finland, and Lithuania. In, Dembner,
S.A. and Perlis, A. (eds), Unasylva – No.198 – Non-Wood Forest Products
and Income Generation. FAO Online Corporate Document Repository.
Available online: http://www.fao.org/docrep/x2450e/x2450e07.htm.
51
Olsson, M.E., Gustavsson, K-E., Andersson, S., Nilsson, A., Duan, R-D. (2004).
Inhibition of Cancer Cell Proliferation in Vitro by Fruit and Berry Extracts
and Correlations with Anti-Oxidant Level. Journal of Agricultural and Food
Chemistry. 52, 7264 – 7271.
Parlane, S., Summers, R.W., Cowie, N.R., Van Gardingen, P.R. (2006).
Management Proposals for Bilberry in Scots Pine Woodland. Forest
Ecology and Management. 222, 272 – 278.
Pengelly, A. (2004). The Constituents of Medicinal Plants. Wallingford, Oxon:
CABI Publishing.
Pliszka, K. (1997). Overview of Vaccinium Production in Europe. In, Yarborogh,
D.E. and Smagula, J.M. (eds.), VII International Symposium on Vaccinium
Culture,ISHS Acta Horticulturae 446. Published Online for the
International Society for Horticultural Science: http://www.actahort.org/.
Prior, R., Guohua, C., Martin, A., Sofic, E., McEwan, J., O’Brien, C., Lischner,
N., Ehlenfeldt, M., Kalt, W., Krewer, G., Mainland, C.M., 1998. Antioxidant
Capacity as Influenced by Total Phenolic and Anthocyanin Content,
Maturity, and Variety of Vaccinium species. Journal of Agricultural and
Food Chemistry. 46, 2686 – 2693.
Rimando, A.M., Kalt, W., Magee, J.B., Dewey, J., Ballington, J.R. (2004).
Resveratrol, Pterostilbene, and Piceatannol in Vaccinium Berries. Journal
of Agricultural and Food Chemistry. 52, 4713 – 4719.
Robinson, Dorienne. (2002). The Powys Wild Food Project. A report
commissioned by Glasu. Available Online:
http://www.glasu.org.uk/reports.htm. Accessed: 24/04/07.
Royal Botanic Garden Edinburgh (2006). Flora Celtica database. RGBE.
Available online:
http://rbg-web2.rbge.org.uk/celtica/dbase/searchformb.html.
Saastamoinen, O. (1999).Forest Policies, Access Rights, and Non-Wood
Forest Products in Northern Europe. In, Dembner, S.A. and Perlis, A.
(eds), Unasylva – No.198 – Non-Wood Forest Products and Income
Generation. FAO Online Corporate Document Repository. Available
online: http://www.fao.org/docrep/x2450e/x2450e06.htm.
Sanderson, H., Prendergast, H.D.V. (2004). Britain’s Wild Harvest: Commercial
Uses of Wild and Traditionally Used Plants in England and Scotland.
London: Kew Publishing in association with The Countryside Agency.
52
Scottish Executive Agricultural and Biological Research Group Flexible Fund.
(2001). Flora Celtica: Sustainable Development of Scottish Plants.
Scottish Executive Central Research Unit. Available online:
http://www.scotland.gov.uk/cru/kd01/orange/sdsp-00.asp.
Selas, V. (2000). Seed Production of a Masting Dwarf Shrub, Vaccinium
myrtillus, in Relation to Previous Reproduction and Weather. Canadian
Journal of Botany. 78, 423 – 429.
Sinclair, F. (2000). Project Blaeberry: A Millennium Award Project. [CDRom]. The Millennium Forest For Scotland Trust. Donated by the author.
Smagula, J.M. (1997). Variation of Fruit Set and Fruit Characteristics of Wild
Lowbush Blueberries (Vaccinium angustifolium Ait.) in a Managed Field.
In, Yarborough, D.E. and Smagula, J.M. (eds.), VI International
Symposium on Vaccinium Culture, ISHS Acta Horticulturae 446.
Published Online for the International Society for Horticultural Science:
http://www.actahort.org/.
Sprieszl, F. (2001). ‘Harvesting the Wild’, An Assessment on [sic] Non-Timber
Forest Products in the Khabarovsk Region of the Russian Far East.
MacGregor Model Forest and the Canadian International Development
Agency. Available Online:
www.mcgregor.bc.ca/downloads/NTFP%20Russia%20Report%20CIDA.p
df. Accessed: 20/08/06.
Stringer B, Kobzik, L. (1998). Environmental Particulate-Mediated Cytokine
Production in Lung Epithelial Cells (A549): Role of Preexisting
Inflammation and Oxidant Stress. Journal of Toxicology and
Environmental Health. 55, 31 – 44.
Talavera, S., Felgines, C., Texier, O., Besson, C., Lamaison, J.L., Remesy,
C. (2003). Anthocyanins are Efficiently Absorbed from the Stomach in
Anesthetized Rats. Journal of Nutrition. 133(12), 4178 – 4182.
Talavera, S., Felgines., C., Texier, O., Besson, C., Manach, C., Lamaison,
J.L., Remesy, C. (2004). Anthocyanins are Efficiently Absorbed from the
Small Intestine. Journal of Nutrition. 134 (9), 2275 – 2279.
Trehane, J. (2004). Royal Horticultural Society Plant Collector Guide,
Blueberries, Cranberries, and Other Vacciniums. Cambridge: Timber
Press.
53
Turnau, K, Mitka, J., Kedzierska, A. (1992). Mycorrhizal Status of Herb-layer
Plants in a Fertilised Oak-Pine Forest. Plant and Soil. 143 (1), 148 – 152.
United States Department of Agriculture Forestry Service, (1999). Alaskan
Special Forest Products Market Report. USDA Forestry Service:
http://www.fs.fed.us/pnw/pubs/gtr500/alaskan.pdf#search=%22mater%20
alaska%20special%20forest%20product%22.
Valentova, K., Ulrichova, J., Cvak, L., Simanek, V. (2007). Cytoprotective
Effect of a Bilberry Extract Against Oxidative Damage of Rat Hepatocytes.
Food Chemistry [online]. Article in press. Available Online:
http://www.sciencedirect.com/science/journal/03088146
Vanderkloet, S.P. (1988). The Genus Vaccinium in North America. Ottawa,
Canada: The Minister of Supply and Services.
Vanderkloet, S.P. and Hill, N.M. (1994). The Paradox of Berry Production in
Temperate Species of Vaccinium. The Canadian Journal of Botany. 72, 52
– 58.
Vickers J. (2005). Getting to the Roots of Soutra’s Old Cures. Available Online:
http://living.scotsman.com/index.cfm?id=1904212005. Accessed 28/03/07.
Wang, H., Guohua, C., Prior, R.L. (1997). Oxygen Radical Abosorbing
Capacity of Anthocyanins. Journal of Agricultural and Food Chemistry. 45,
304 – 309.
Witzell J., Gref R., Näsholm T. (2003). Plant-part Specific and Temporal
Variation in Phenolic Compounds of Boreal Bilberry (Vaccinium myrtillus)
Plants. Biochemical Systematics and Ecology. 31, 115 – 127.
Yarborough, D.E. (1997). Production Trends in the wild Blue Berry Industry in
North America. In, Yarborough, D.E. and Smagula, J.M. (eds.), VI
International Symposium on Vaccinium Culture, ISHS Acta Horticulturae
446. Published Online for the International Society for Horticultural
Science: http://www.actahort.org/.
Yarborough, D.E. (2002). Development of a Crop Estimation Technique for Wild
Blueberries. In, Hepp, R.F. (ed.), VII International Symposium on
Vaccinium culture, ISHS Acta Horticulturae 574. Published Online for the
International Society for Horticultural Science: http://www.actahort.org/.
54
Yarborough, D.E. (2002). Progress Towards the Development of a Mechanical
Harvester for Wild Blueberries. In, Hepp, R.F. (ed.), VII International
Symposium on Vaccinium culture, ISHS Acta Horticulturae 574. Published
Online for the International Society for Horticultural Science:
http://www.actahort.org/.
Yi, W., Fischer, J., Krewer, G., Akoh, C.C. (2005). Phenolic Compounds from
Blueberries Can Inhibit Colon Cancer Cell Proliferation and Induce
Apoptosis. Journal of Agricultural and Food Chemistry. 53(18), 7320 –
7329.
Zadernowski, R., Naczk, M., Nesterowicz, J. (2005). Phenolic Acid Profiles in
Some Small Berries. Journal of Agricultural and Food Chemistry. 53(6),
2118 – 2124.
Zhang, Z., Xiaolan, K., Fugal, K., McLaughlin, J. (2004). Comparison of HPLC
Methods for Determination of Anthocyanins and Anthocyanidins in Bilberry
Extract. Journal of Agricultural and Food Chemistry. 52, 688 – 691.
Zhao, C., Giusti, M.M., Malik, M., Moyer, M.P., Magnuson, B.A. (2004).
Effects of Commercial Anthocyanin-Rich Extracts on Colonic Cancer and
Nontumorigenic Colonic Cell Growth. Journal of Agricultural and Food
Chemistry. 52, 6122 – 6128.
55
Appendix One: Bibliography
Albert T, Raspe, O., Jaquemart A-L. (2004). Clonal Diversity and Genetic
Structure in Vaccinium myrtillus Populations from Different Habitats.
Belgian Journal of Botany. 137 (2), 155 – 162.
Allen, D.E., and Hatfield, G. (2004). Medicinal Plants in Folk Tradition: An
Ethnobotany of Britain and Ireland. Cambridge: Timber Press.
Australian Government Rural Industries Research and Development
Corporation (2006). Medicinal Herbs – A Preliminary Evaluation of
Astragalus, Bilberry, Feverfew, and Stevia. Tazmania: AGRIRDC,
(05/185).
Beccaro, G., Mellano, M.G., Chiabrando, V., Bounous, G. (2006). Phenolic and
Anthocanin Content and Antioxidant Activity in Fruits of Bilberry
(Vaccinium myrtillus L.) and of Highbush Blueberry (V. corymbosum L.)
Cultivars in North Western Italy. International Society for Horticultural
Science: Proceedings of the VIIIth International Symposium on Vaccinium
Culture. Acta Horticulturae. 715, 553 - 557. Available Online to
subscribers: http://www.ishs.org/acta/index.htm.
Beith, M. (1995). Healing Threads: Traditional Medicines of the Highlands and
Islands. Edinburgh: Birlinn, Limited
Biffignandi, P.M., and Carletto, L. (2000). The New Attitude of the European
Regulatory Authorities about Herbal Medicinal Products. Drug Information
Journal. 34, 801 – 808.
Boulakia, F.C., Valensi, P.E., Boulahdour, H., Lestrade, R., Dufour-Lamartinie, JF., Hort-Legrand, C., Behar, A. (2000). In Vivo Sequential Study of
Skeletal Muscle Capillary Permeability in Diabetic Rats: Effect of
Anthocyanosides. Metabolism. 49 (7), 880 – 885.
Brielmann, H.L. (1999). Phytochemicals: The Chemical Components of Plants.
In, Kaufmann, P.B., Cseke, L.J., Warber, S., Duke, J.A., Brielmann, H.L.,
Natural Products from Plants. London: CRC Press LLC.
56
Brown, D.M., Donaldson, K., Borm, P.J., Schins, R.P., Dehnhardt, M., Gilmour,
P., Jimenez, L.A., Stone, V. (2004). Calcium and ROS-meditated
Activation of Transcription Factors and TNF-alpha Cytokine Gene
Expression in Macrophages Exposed to Ultrafine Particles. American
Journal of Physiology, Lung Cellualr and Molecular Physiology. 286, L344
– L353.
Bruneton, J. (1999). Pharmacognosy: Phytochemistry Medicinal Plants. 2nd
ed. Paris: Lavoisier Publishing.
Canadian International Development Agency. (2001). “Harvesting the Wild”:
An Assessment of Non-Timber Forest Products in the Khaborovsk Region
of the Russian Far East. McGregor Model Forest Association. Available
online: http://www.mcgregor.bc.ca.
Canter, P. and Ernst, E. (2004). Anthocyanins of Vaccinium myrtillus (Bilberry)
For Night Vision – A Systematic Review of Placebo-Controlled Trials.
Survey of Opthamology. 49 (1), 38 – 50.
Cao, G., Muccitelli, Sanchez – Moreno, C., Prior, R. (2001). Anthocyanins
Are Absorbed in Glycated Forms In Elderly Women: A Pharmacokinetic
Study. American Journal of Clinical Nutrition. 73, 920 – 926.
Chatterjee, A., Yasmin, T., Bagchi, D., Stohs, S.J. (2004). Inhibition of
Helicobacter pylori in vitro by various berry extracts, with enhanced
susceptibility to clarithromycin. Molecular and Cellular Biochemistry.
265 (1- 2), 19 – 26.
Chrimes, D. (2004). Stand Development and Regeneration Dynamics of
Managed Uneven-Aged Picea abies Forests in Boreal Sweden. Thesis
(PhD). Swedish University of Agricultural Sciences.
Cignarella, A., Nastasi M., Cavalli E., Puglisi, L. (1996). Novel Lipid-Lowering
Properties of Vaccinium myrtillus L. Leaves, A Traditional Antidiabetic
Treatment, In Several Models of Rat Dyslipidaemia: A Comparison with
Ciprofibrate. Thrombosis Research. 84 (5), 311 – 322.
Cline, W.O. (1997). Postharvest Infection of Blueberries During Handling. In,
Yarborough, D.E. and Smagula, J.M. (eds.), VI International Symposium
on Vaccinium Culture, ISHS Acta Horticulturae 446. Published Online for
the International Society for Horticultural Science:
http://www.actahort.org/.
57
Cohen – Boulakia, F., Valensi, P.E., Boulahdour, H., Lestrade, R., Doufour –
Lamartinie, J.F., Hort – Legrande, C., Behar, A. (2000). In Vivo Sequential
Study of Skeletal Muscle Capillary Permeability in Diabetic Rats: Effect of
Anthocyanosides. Metabolism. 49 (7): 880 – 5.
Comhairle nan Eileen Siar. (2007). Liostaichean. Available Online:
http://www.cne-siar.gov.uk/gaelic/grd/liostaichean/lusan/lusan_f.htm.
Accessed 28/03/07.
Cooke, D., Steward, W.P., Gescher, A.J., Marczylo, T. (2005). Anthocyans from
Fruits and Vegetables – Does Bright Colour Signal Cancer
Chemoprotective Activity? European Journal of Cancer. 41, 1931 – 1940.
Craig, W. (1879). Milne’s Manual of Materia Medica and Therapeutics.
Edinburgh: E. & S. Livingstone.
Culpeper, J. (1981 (1826)). Culpeper’s Complete Herbal & English Physician.
Barcelona: Harvey Sales.
Dewick, P.M. (2001). Medicinal Natural Products, A Biosynthetic Approach.
Chichester: John Wiley & Sons Ltd.
Dickson C., and Dickson, J. (2000). Plants and People in Ancient Scotland.
Gloucestershire: Tempus Publishing.
Dickson, J.H., Oeggl, K., Holden T.G., Handley, L.L., O’Connell, T.C.,
Preston, T. (2000). The Omnivorous Tyrolean Iceman: Colon Contents
(meat, cereals, pollen, moss, and whipworm) and Stable Isotope Analysis.
Philosophical Transactions of the Royal Society London B. 355, 1843 –
1849.
Donaldson, K., Stone, V., Borm, P.J., Jimenez, L.A., Gilmour, P.S., Schins, R.P.,
Knaapen, A.M., Rahman, I., Faux, S.P., Brown, D.M., MacNee, W. (2003).
Oxidative Stress and Calcium Signaling in the Adverse Effects of
Environmental Particles (PM10). Free Radical Biology & Medicine. 34
(11), 1369 – 1382.
Du, Q., Jerz, G., Winterhalter, P. (2004). Isolation of Two Anthocyanin
Sambubiosides from Bilberry (Vaccinium myrtillus) by High-Speed
Counter-Current Chromatography. Journal of Chromatography A.
1045, 59 – 63.
Dugo, P., Mondello, L., Errante, G., Zappia, G., Dugo, G. (2001). Identification
of Anthocyanins in Berries by Narrow-Bore High-Performance Liquid
Chromatography with Electrospray Ionisation Detection. Journal of
Agricultural and Food Chemistry. 49, 3987 – 3992.
58
Duncan, A. (1829). Supplement to the Edinburgh New Dispensary. Edinburgh:
Bell & Bradfute.
Ehala, S., Vaher, M., Kaljurand, M. (2005). Characterization of Phenolic
Profiles of Northern European Berries by Capillary Electrophosphoresis
And Determination of their Antioxidant Activity. Journal of Agricultural and
Food Chemistry. 53 (16), 6484 – 6490.
Ehlenfeldt, M.K. (2002). Postharvest Research and Technology in Vaccinium.
In, Hepp, R.F. (ed.), VII International Symposium on Vaccinium culture,
ISHS Acta Horticulturae 574. Published Online for the International
Society for Horticultural Science: http://www.actahort.org/.
Emerit, J., Beaumont, C., Trivin, F. (2001). Iron Metabolism, Free Radicals, and
Oxidative Injury. Biomedical Pharmacotherapy. 55 (6), 333 – 339.
Emery, M., Martin, S., Dyke, A. (2006). Wild Harvests from Scottish Woodlands,
Social, Cultural, and Economic Values of Contemporary Non-Timber
Forest Products. Edinburgh: Forestry Commission. Available Online:
http://www.forestreasearch.gov.uk/fr/INFD-6XCHWF.
Erlund, I. (2002). Chemical Analysis and Pharmacokinetics of the Flavanoids
Quercetin, Hesperetin, and Naringenin in Humans. Thesis (PhD).
University of Helsinki.
Ernst, E. (2000). Herbal Medicines: Where is the Evidence? British Medical
Journal. 321, 395 – 396.
Evans, W.C. (2002). Trease and Evans Pharmacology (15th ed). Edinburgh:
W.B. Saunders Company Limited.
Faria, A., Calhau, C., de Freitas, V., Mateus, N. (2006). Procyanidins as
Antioxidants and Tumor Cell Growth Modulators. Journal of Agricultural
and Food Chemistry. 54 (6) – 2392 – 2397.
Faria, A., Oliveira, J., Neves, P., Gameiro, P., Santos – Buelga, C., de Freitas,
V., Mateus, N. (2005). Antioxidant Properties of Prepared Blueberry
(Vaccinium myrtillus) Extracts. Journal of Agricultural and Food Chemistry.
53(17), 6896 – 6902.
Featherstone, A.W. (2002). Blaeberry (Vaccinium myrtillus) Species Profile.
Caledonian Forest Species Profiles. Published by Caledonia Wild!
Available Online: http://www.treesforlife.org.uk/tfl.blaeberry.html.
Accessed: 15/04/07
59
Finn, C. 1(999). Temperate Berry Crops. In, Janick, J. (ed), Perspectives on
New Crops and New Uses. Alexandria, VA: ASHS Press.
Grieve, Maud. (1971). A Modern Herbal: Vol. I, A – H. New York: Dover
Publications, Inc.
Guohua, C., Muccitelli, H.U., Sanchez-Moreno, C., Prior, R. (2001).
Anthocyanins are Absorbed in Glycated Forms in Elderly Women: A
Pharmakokinetic Study. American Journal of Clinical Nutrition. 73, 920 –
926.
Honkavaara, J., Siitari, H., Saloranta, V., Vitala, J. (2007). Avian Seed Ingestion
Changes Germination Patterns of Bilberry, Vaccinium myrtillus. Annales
Botanici Fennici. 44, 8 – 17.
Harborne, J.B. (1973). Phytochemical Methods: A Guide to Modern Techniques
of Plant Analysis. 2nd ed. London: Chapman and Hall
He, J., Magnuson, B.A., Giusti, M.M. (2005). Analysis of Anthocyanins in Rat
Intestinal Contents – Impact of Anthocyanin Chemical Structure on Fecal
Excretion. Journal of Agricultural and Food Chemistry. 53, 2859 – 2866.
Hinckleton, P.R., Reekie, J.Y.C., MacKenzie, K., Eaton, L.J., Havard, P. (2002).
Freeze Damage and Frost Tolerance Thresholds for Flowers of the
Lowbush Blueberry (Vaccinium angustifolium Ait.). In, Hepp, R.F. (ed.), VII
International Symposium on Vaccinium culture, ISHS Acta Horticulturae
574. Published Online for the International Society for Horticultural
Science: http://www.actahort.org/.
Hou, D-X., Yanagita, T., Uto, T., Masuzaki, S., Fujii, M. (2005). Anthocyanidins
Inhibit Cyclooxygenase-2 Expression in LPS-evoked Macrophages:
Structure-Activity Relationship and Molecular Mechanisms Involved.
Biochemical Pharmacology. 70, 417 – 425.
Ihalainen, M., Alho, J., Kohlehmainen, O., Pukkala, T. (2002). Expert Models
for Bilberry and Cowberry Yields in Finnish Forests. Forest Ecology and
Management. 157, 15 – 22.
Jaakola, L. (2003). Flavonoid Biosynthesis in Bilberry (Vaccinium myrtillus L.).
Thesis (Ph.D.). University of Oulu.
Jayaprakasam, B., Vareed, S.K., Olsen, L.K., Nair, M.G. (2005). Insulin
Secretion by Bioactive Anthocyanins and Anthocyanidins present in Fruits.
Journal of Agricultural and Food Chemistry. 53 (1), 28 – 31
60
Jensen, H.D., Krogfelt, K.A, Cornett, C., Hansen, S.H., Christensen, S.B.
(2002). Hydrophilic Carboxylic Acids and Iridoid Glycosides in the Juice of
American and European Cranberries (Vaccinium macrocarpon and V.
oxycoccus), Lingonberries (V. vitis-idaea), and Blueberries. Journal of
Agricultural and Food Chemistry. 50, 6871 – 6874.
Johnson, C.P. (1862). A Treatise upon the Principle Native Vegetables
Capable of Application as Food, Medicine, or in the Arts and
Manufactures. London: Robert Hardwicke.
Kahkonen, M.P., Heinamaki, J., Ollilainen, V., Heinonin, M. (2003). Berry
Anthocyanins: Isolation, Identification, and Antioxidant Activities. Journal
of the Science of Food and Agriculture. 83, 1403 – 1411.
Kang, S-Y., Seeram, N., Nair, M.G., Bourquin, L.D. (2003). Tart Cherry
Anthocyanins Inhibit Tumor Development in APCMin Mice and Reduce
Prolferation of Human Colon Cancer Cells. Cancer Letters. 194, 13 – 19.
Kangas, K. (1999). Trade of Main Wild Berries in Finland. Silva Fennica. 33
(2), 159 – 168.
Katsube, N., Iwashita, K., Tsushida, T., Yamaki, K., Kobori. M. (2003).
Induction of Apoptosis in Cancer Cells by Bilberry (Vaccinium myrtillus)
and the Anthocyanins. Journal of Agricultural and Food Chemistry. 51 (1),
68 – 75.
Kay, C. (2006). Aspects of Anthocyanin Absorption, Metabolism, and
Pharmacokinetics in Humans. Nutrition Research Reviews. 19, 137 – 146.
Kay, C. and Holub, B.J. (2002). The Effect of Wild Blueberry (Vaccinium
angustifolium) Consumption on Postprandial Serum Antioxidant Status in
Human Subjects. British Journal of Nutrition. 88, 389 – 397.
Kay, C., Mazza, G., Holub, B.J., Wang, J. (2004). Anthocyanin Metabolites in
Human Urine and Serum. British Journal of Nutrition. 91, 933 – 942.
King, J.W., Grabiel, R.D., and Wightman, J.D. (date unknown). Subcritical Water
Extraction of Anthocyanins from Fruit Berry Substrates. Available Online:
scrub.lanl.gov/pdf/king/192_subcritical_water.pdf. Accessed: 27/04/07.
Koenig, W. and Arnold, D. (1961). Facsimile of the First Amsterdam
Pharmacopoaeia, 1636. Nieuwkoop: B. De Graaf.
Kolosova, N.G., Shcheglova, T.V., Sergeeva, S.V., Loskutova, L.V. (2006)
Long-Term Anti-oxidant Supplementation Attenuates Oxidative Stress
Markers and Cognitive Deficits in Senescent-Accelerated OXYS Rats.
Neurobiology of Aging. 27(9), 1289 – 1297.
61
Konovalchuk V.K. and Konovalchuk V.V. (2006) The Resources of Wild Lowbush
Blueberries (Vaccinium myrtillus L., Vaccinium uglinosum L.) and
Highbush Blueberry culture in Ukraine. In, Lopes da Fonseca, F., Romero
Munoz, F. (eds.), VIII International Symposium on Vaccinium Culture,
ISHS Acta Horticulturae 715. Published Online for the International
Society for Horticultural Science: http://www.actahort.org/.
Korkina, L.G. and Afanas’ev, I.B. (1997). Antioxidant and Chelating Properties of
Flavonoids. In, Sies, H. (ed.), Antioxidants in Disease Mechanisms and
Therapy. In, August, J.T., Murad, F., Anders, M.W., and Coyle, J.T. (eds.),
Advances in Pharmacology, Volume 38. London: Academic Press.
Lala, G. (2005). Effects of Anthocyanin Rich Extracts on Multiple Biomarkers
of Colon Cancer. Thesis (M.Sc.). University of Maryland, College Park.
Lang, D.C. (1987). The complete Book of British Berries. London: Threshold
Books.
Leonard, H.C. (1892). Dictionary of Materia Medica and Therapeutics.
London: Bailliere, Tindall, and Cox.
Leven, E. Segerstedt, A. (2004). Polarica’s Wild Berries: An Example of a
Required Storage Capacity Calculation and Where to Locate this
Inventory. Supply Chain Management. 9(3), 213 – 218.
Lietti, A., Cristoni, A., Picci, M. (1976). Studies of Vaccinium myrtillus
Anthocyanosides I: Vasoprotective and Antiinflammatory Activity.
Arzneimittelforschung. 26(5), 829 – 832.
Lietti, A., Forni, G. (1976). Studies on Vaccinium myrtillus anthocyanosides II:
Aspects of Anthocyanin Pharmacokinetics in the Rat.
Arzneimittelforschung. 26(5), 832 – 835.
Lila, M. A. (2004). Anthocyanins and Human Health: An Investigative
Approach. Journal of Biomedicine and Biotechnology. 5, 306 – 313.
Logan, A.C. and Wong, C. (2001). Chronic Fatigue Syndrome: Oxidative Stress
and Dietary Modifications. Alternative Medicine Review. 6(5), 450 –
459.
Lyons, M.M., Yu, C., Toma, R.B., Cho, S.Y., Reiboldt, W., Lee, J., Van
Breemen, R.B. (2003). Resveratrol in Raw and Baked Blueberries and
Bilberries. Journal of Agricultural and Food Chemistry. 51, 5867 – 5870.
62
MacBain, Alexander. (1982). An Etymological Dictionary of the Gaelic Language.
Glasgow: Gairm Publications. Available Online:
http://www.ceantar.org/Dicts/MB2/index.html. Accessed: 28/03/07.
MacKay, J.W.B. (2004). Effect of Burning on the Mycorrhizal fungi of Scots Pine
Pinus sylvestris at Boat of Garten, Inverness-shire, Scotland.
Conservation Evidence. 1, 20 – 22. Available Online:
www.conservationevidence.com/Attachments/PDF83.pdf. Accessed:
20/04/07.
Madhavi D.L., Bomser J., Smith M.A.L., Singletary K. (1998). Isolation of
Bioactive Constituents from Vaccinium myrtillus (bilberry) Fruits and Cell
Cultures. Plant Science. 131, 95 – 103.
Mills, S and Bone, K. (2000). Principles and Practice of Phytotherapy, Modern
Herbal Medicine. 1st ed. Edinburgh: Churchill Livingstone.
Moffatt, B. and Fulton, J. (1988). SHARP Practice 2: The Second Report on
Researches into the Medieval Hospital at Soutra, Lothian Region,
Scotland. Edinburgh: Soutra Hospital Archaeoethnopharmacological
Project.
Moffatt. B, Thompson, B., Fulton, J. (1989). SHARP Practice 3: The Third
Report on Researches into the Medieval Hospital at Soutra, Lothian
Region, Scotland. Edinburgh: Soutra Hospital
Archaeoethnopharmacological Project.
Molina, R, Vance., N., Weigand, J.F., Pilz, D., Amaranthus, M.P. (1997).
Special Forest Products: Integrating Social, Economic, and Biological
Considerations into Ecosystem Management. Available online:
http://www.fs.fed.us/pnw/pubs/journals/pnw_1997_molina001.pdf.
Morazzoni P., Bombardelli, E. (1996). Vaccinium Myrtillus L.. Fitoterapia. 61 (1),
3 – 29).
Moyer, RA., Hummer, K.E., Finn C.E., Frei, B., Wrolstad, R.E. (2002).
Anthocyanins, Phenolics, and Antioxidant Capacity in Diverse Small
Fruits: Vaccinium, Rubus, and Ribes. Journal of Agricultural and Food
Chemistry. 50, 519 – 525.
Murray, J. (1825). A System of Materia Medica and Pharmacy, Vol. II.
Edinburgh: Adam Black.
Muth, E.R., Laurent, J.M., Jasper, P., 2000. The Effect of Bilberry Nutritional
Supplementation on Night Visual Acuity and Contrast Sensitivity.
Alternative Medicine Review. 5 (2), 164 – 173.
63
Nakajima, J-I., Tanaka, I., Seo, S., Mamazaki, M., Saito, K. (2004).
LC/PDA/MS Profiling and Radical Scavenging Activity of Anthocyanins in
Various Berries. Journal of Biomedicine and Biotechnology. 2004(5), 241
– 247.
[No Authors Listed] (2001). Monograph: Vaccinium myrtillus (bilberry).
Alternative Medicine Review. 6(5), 450 – 459.
Nuortila C., Tuomi, J., And Laine, K. (2002). Inter-parent Distance Affects
Reproductive Success in Two Clonal Dwarf Shrubs, Vaccinium myrtillus
and Vaccinium vitis-idaea (Ericaceae). Canadian Journal of Botany. 80,
875 – 884.
Nyman, N.A., Kumpulainen, J.T. (2001). Determination of Anthocyanins in
Berries and Red Wine by High-Performance Liquid Chromatography.
Journal of Agricultural and Food Chemistry. 49(9), 4183 – 4187.
Olmos, S. (1999). Non-Wood Forest Products: Utilization and Income
Generation in the Czech Republic, Finland, and Lithuania. In, Dembner,
S.A. and Perlis, A. (eds), Unasylva – No.198 – Non-Wood Forest Products
and Income Generation. FAO Online Corporate Document Repository.
Available online: http://www.fao.org/docrep/x2450e/x2450e07.htm.
Olsson, M.E., Gustavsson, K-E., Andersson, S., Nilsson, A., Duan, R-D. (2004).
Inhibition of Cancer Cell Proliferation in Vitro by Fruit and Berry Extracts
and Correlations with Anti-Oxidant Level. Journal of Agricultural and Food
Chemistry. 52, 7264 – 7271.
Pap, N., Pongracz, E., Myllykoski, L., Keiski, R., (2004). Waste Minimization
and Utilization in the Food Industry: Processing of Artic Berries, and
Extraction of Valuable Compounds from Juice-Processing By-Products. In,
Pongracz, E. (ed), Proceedings of the Waste Minimization and Resources
Use Optimization Conference. Oulu, Finland: Oulu University Press, 159 –
168.
Parlane, S., Summers, R.W., Cowie, N.R., Van Gardingen, P.R. (2006).
Management Proposals for Bilberry in Scots Pine Woodland. Forest
Ecology and Management. 222, 272 – 278.
Pengelly, A. (2004). The Constituents of Medicinal Plants. Wallingford, Oxon:
CABI Publishing.
64
Percival, D., Crowe, N., Stevens, D., Laughlin, P.O. (2002). Reduction in
Anthocyanin Exudation from Individual Quick Frozen Lowbush Blueberries
(Vaccinium angustifolium Ait.) Through Use of Edible Films. . In, Hepp,
R.F. (ed.), VII International Symposium on Vaccinium culture, ISHS Acta
Horticulturae 574. Published Online for the International Society for
Horticultural Science: http://www.actahort.org/.
Pliszka, K. (1997). Overview of Vaccinium Production in Europe. In, Yarborough,
D.E. and Smagula, J.M. (eds.), Sixth International Symposium on
Vaccinium Culture, ISHS Acta Horticulturae 446. Published Online for the
International Society for Horticultural Science: http://www.actahort.org/.
Prendergrast H.D.W., and Sanderson, H. (2004). Britain’s Wild Harvest:
Commercial Uses of Wild Plants and Fungi. China: Compass Press Ltd.,
for Royal Botanic Garden Kew.
Prior, R., Guohua, C., Martin, A., Sofic, E., McEwan, J., O’Brien, C., Lischner,
N., Ehlenfeldt, M., Kalt, W., Krewer, G., Mainland, C.M., 1998. Antioxidant
Capacity as Influenced by Total Phenolic and Anthocyanin Content,
Maturity, and Variety of Vaccinium species. Journal of Agricultural and
Food Chemistry. 46, 2686 – 2693.
Rimando, A.M., Kalt, W., Magee, J.B., Dewey, J., Ballington, J.R. (2004).
Resveratrol, Pterostilbene, and Piceatannol in Vaccinium Berries. Journal
of Agricultural and Food Chemistry. 52, 4713 – 4719.
Robinson, Dorienne. (2002). The Powys Wild Food Project. A report
commissioned by Glasu. Available Online:
http://www.glasu.org.uk/reports.htm. Accessed: 24/04/07.
Royal Botanic Garden Edinburgh (2006). Flora Celtica database. RGBE.
Available online:
http://rbg-web2.rbge.org.uk/celtica/dbase/searchformb.html.
Selas, V. (2000). Seed Production of a Masting Dwarf Shrub, Vaccinium
myrtillus, in Relation to Previous Reproduction and Weather. Canadian
Journal of Botany. 78, 423 – 429.
Sanderson, H., Prendergast, H.D.V. (2004). Britain’s Wild Harvest: Commercial
Uses of Wild and Traditionally Used Plants in England and Scotland.
London: Kew Publishing in association with The Countryside Agency.
65
Saastamoinen, O. (1999).Forest Policies, Access Rights, and Non-Wood
Forest Products in Northern Europe. In, Dembner, S.A. and Perlis, A.
(eds), Unasylva – No.198 – Non-Wood Forest Products and Income
Generation. FAO Online Corporate Document Repository. Available
online: http://www.fao.org/docrep/x2450e/x2450e06.htm.
Schmidt, B.M., Erdman, J.W., Lila, M.A. (2005). Effects of Food Processing on
Blueberry Antiproliferation and Antioxidant Activity. Journal of Food
Science. 70, s389 – s394.
Scottish Executive Agricultural and Biological Research Group Flexible Fund.
(2001). Flora Celtica: Sustainable Development of Scottish Plants.
Scottish Executive Central Research Unit. Available online:
http://www.scotland.gov.uk/cru/kd01/orange/sdsp-00.asp.
Silano, M., De Vincenzi, M., De Vincenzi, A., Silano, V. (2004). The New
European Legislation on Traditional Herbal Medicines: Main Features and
Perspectives. Fitoterapia. 75, 107 – 116.
Sinclair, F. (2000). Project Blaeberry: A Millennium Award Project. [CDRom]. The Millennium Forest for Scotland Trust. Donated by the author.
Sinitsina, O., Krysanova, Z., Ishchenko, A., Dikalova, A.E., Stolyarov, S.,
Kolosova, N. (2006). Age-Associated Changes in Oxidative Damage and
the Activity of Antioxidant Enzymes in Rats with Inherited Overgeneration
of Free Radicals. Journal of Cellular and Molecular Medicine. 10 (1), 206 –
215.
Smagula, J.M. (1997). Variation of Fruit Set and Fruit Characteristics of Wild
Lowbush Blueberries (Vaccinium angustifolium Ait.) in a Managed Field.
In, Yarborough, D.E. and Smagula, J.M. (eds.), VI International
Symposium on Vaccinium Culture, ISHS Acta Horticulturae 446.
Published Online for the International Society for Horticultural Science:
http://www.actahort.org/.
Sprieszl, F. (2001). ‘Harvesting the Wild’, An Assessment on [sic] Non-Timber
Forest Products in the Khabarovsk Region of the Russian Far East.
MacGregor Model Forest and the Canadian International Development
Agency. Available Online:
www.mcgregor.bc.ca/downloads/NTFP%20Russia%20Report%20CIDA.p
df. Accessed: 20/08/06.
66
Stringer B, Kobzik, L. (1998). Environmental Particulate-Mediated Cytokine
Production in Lung Epithelial Cells (A549): Role of Preexisting
Inflammation and Oxidant Stress. Journal of Toxicology and
Environmental Health. 55, 31 – 44.
Talavera, S., Felgines, C., Texier, O., Besson, C., Lamaison, J.L., Remesy,
C. (2003). Anthocyanins are Efficiently Absorbed from the Stomach in
Anesthetized Rats. Journal of Nutrition. 133(12), 4178 – 4182.
Talavera, S., Felgines., C., Texier, O., Besson, C., Manach, C., Lamaison,
J.L., Remesy, C. (2004). Anthocyanins are Efficiently Absorbed from the
Small Intestine. Journal of Nutrition. 134 (9), 2275 – 2279.
Trehane, J. (2004). Royal Horticultural Society Plant Collector Guide,
Blueberries, Cranberries, and Other Vacciniums. Cambridge: Timber
Press.
Turnau, K, Mitka, J., Kedzierska, A. (1992). Mycorrhizal Status of Herb-layer
Plants in a Fertilised Oak-Pine Forest. Plant and Soil. 143 (1), 148 – 152.
United States Department of Agriculture Forestry Service, (1999). Alaskan
Special Forest Products Market Report. USDA Forestry Service:
http://www.fs.fed.us/pnw/pubs/gtr500/alaskan.pdf#search=%22mater%20
alaska%20special%20forest%20product%22.
Valentova, K., Ulrichova, J., Cvak, L., Simanek, V. (2007). Cytoprotective
Effect of a Bilberry Extract Against Oxidative Damage of Rat Hepatocytes.
Food Chemistry [online]. Article in press. Available from:
http://www.sciencedirect.com/science/journal/03088146.
Vanderkloet, S.P. (1988). The Genus Vaccinium in North America. Ottawa,
Canada: The Minister of Supply and Services.
Vanderkloet, S.P. and Hill, N.M. (1994). The Paradox of Berry Production in
Temperate Species of Vaccinium. The Canadian Journal of Botany. 72, 52
– 58.
Vickers J. (2005). Getting to the Roots of Soutra’s Old Cures. Available Online:
http://living.scotsman.com/index.cfm?id=1904212005. Accessed 28/03/07.
Viljanen, K. (2005). Protein Oxidation and Protein-Lipid Interactives in
Different Food Models in the Presence of Berry Phenolics. Thesis (PhD).
University of Helsinki.
67
Viljanen, K., Kylli, P., Kivikari, R., Heinonen, M. (2004). Inhibition of protein
and Lipid Oxidation in Liposomes by Berry Phenolics. Journal of
Agricultural and Food Chemistry. 52(24), 7419 – 7424.
Wang, H., Guohua, C., Prior, R.L. (1997). Oxygen Radical Abosorbing
Capacity of Anthocyanins. Journal of Agricultural and Food Chemistry. 45,
304 – 309.
Witzell J., Gref R., Näsholm T. (2003). Plant-part Specific and Temporal
Variation in Phenolic Compounds of Boreal Bilberry (Vaccinium myrtillus)
Plants. Biochemical Systematics and Ecology. 31, 115 – 127
Yarborough, D.E. (1997). Production Trends in the Wild Blueberry Industry in
North America. In, Yarborough, D.E. and Smagula, J.M. (eds.), VI
International Symposium on Vaccinium Culture, ISHS Acta Horticulturae
446. Published Online for the International Society for Horticultural
Science: http://www.actahort.org/.
Yarborough, D.E. (2002). Development of a Crop Estimation Technique for Wild
Blueberries. In, Hepp, R.F. (ed.), VII International Symposium on
Vaccinium culture, ISHS Acta Horticulturae 574. Published Online for the
International Society for Horticultural Science: http://www.actahort.org/.
Yarborough D.E. and Marra, M.C. (1997). Economic Threshholds for Weeds in
Wild Blueberry fields. In, Yarborough, D.E. and Smagula, J.M. (eds.), VI
International Symposium on Vaccinium Culture, ISHS Acta Horticulturae
446. Published Online for the International Society for Horticultural
Science: http://www.actahort.org/.
Yarborough, D.E. (2002). Progress Towards the Development of a Mechanical
Harvester for Wild Blueberries. In, Hepp, R.F. (ed.), VII International
Symposium on Vaccinium culture, ISHS Acta Horticulturae 574. Published
Online for the International Society for Horticultural Science:
http://www.actahort.org/.
Yi, W., Fischer, J., Krewer, G., Akoh, C.C. (2005). Phenolic Compounds from
Blueberries Can Inhibit Colon Cancer Cell Proliferation and Induce
Apoptosis. Journal of Agricultural and Food Chemistry. 53(18), 7320 –
7329.
Zadernowski, R., Naczk, M., Nesterowicz, J. (2005). Phenolic Acid Profiles in
Some Small Berries. Journal of Agricultural and Food Chemistry. 53(6),
2118 – 2124.
68
Zhang, H.L., Bushway, A.A., Work, T., Camire, M.E., Work, R. (1997). Prevention
of Anthocyanin Leakage of Individually Quick Frozen (IQF) Lowbush
Blueberries in Blueberry Muffins. In, Yarborough, D.E. and Smagula, J.M.
(eds.), VI International Symposium on Vaccinium Culture, ISHS Acta
Horticulturae 446. Published Online for the International Society for
Horticultural Science: http://www.actahort.org/.
Zhang, Z., Xiaolan, K., Fugal, K., McLaughlin, J. (2004). Comparison of HPLC
Methods for Determination of Anthocyanins and Anthocyanidins in Bilberry
Extract. Journal of Agricultural and Food Chemistry. 52, 688 – 691.
Zhao, C., Giusti, M.M., Malik, M., Moyer, M.P., Magnuson, B.A. (2004).
Effects of Commercial Anthocyanin-Rich Extracts on Colonic Cancer and
Nontumorigenic Colonic Cell Growth. Journal of Agricultural and Food
Chemistry. 52, 6122 – 6128.
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Appendix Two: Blaeberry Reproduction and Mycorrhizal Dependence
A 1994 study of Canadian soil samples by Vaccinium expert S.P.
Vanderkloet found that seeds from this genus were underrepresented in the seed
banks of soils where they are found in North America (Vandekloet and Hill 1994,
56) even where the plants were growing abundantly. He hypothesized that loss
of seeds may be taking place through dispersal by birds, between seed
deposition and establishment in the soil, through germination, or because of
fungal decomposition (ibid.). Contrary to earlier studies (as cited in Vanderkloet
and Hill 1994, 57), a recently published study by Honkavaara et. al. (2007) found
an increase in rates of germination for Vaccinium myrtillus seeds when they were
passed through the digestive systems of thrushes (Turdus spp.). They found that
germination rates increased for ‘passed’ seeds except those from berries picked
very early in the season (Honkavaara et.al. 2007, 15). Variation in seed
germination rates also occurred between sample-years, leading them to
hypothesize that that there is potential variation in germination success within the
berry season itself, between crop years, and when seeds are eaten by birds or
foragers (ibid., 15 – 16).
Since sexual reproduction is known to not be particularly successful in
Vaccinium myrtillus, research is also focusing on clonal reproduction. Blaeberry
plants have a rhizome system from which new shoots may arise for up to 15
years (Featherstone 2002). This system also allows the plants to survive burning
episodes, as may be the case in forest fires or crop-maintenance practices.
Rhizomatous propagation may also be used commercially as a means of rapidly
generating fruit-bearing plants. However, as with all clonal plants, genetic
diversity and plant reproductive health must be considered. A recent study of
clonal diversity in Belgium (Albert et. al. 2004) took samples of blaeberry from 3 x
3 m plots across a population and sampled them using RAPD procedure. They
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found that genetic diversity within a local population was highly variable and that
Vaccinium myrtillus seems to use a phalanx growth structure in which ramets
belonging to a clone are grouped closely together to form clumps (Albert et. al.
2004, 158). They attribute the genetic variation to a number of factors including
the ‘architectural parameters’ of rhizome formation (e.g. internode length and
branching angle) and ‘environmental heterogeneity’ and they hypothesize that
sexual reproduction within a population can only take place within windows of
opportunity, explaining why seedlings and seed banks are often missing from
Vaccinium populations (ibid. 160).
Nuortila et. al. (2002) also examined reproductive success in clonal
Vaccinium myrtillus plants by examining the pollination patterns of closely
situated plants both by hand and through pollination by Bombus spp. Their
hypothesis, that proximal plants would be closely genetically related and that this
might lower seed production due to inbreeding or self-incompatibility, was found
to have foundation. They found that in northern and southern boreal forests of
Finland pollination was entirely done by bees, whereas in other experiments in
Belgium found a 10% fruit set through self-pollination alone (Nuortila et. al. 881).
Additionally, they found that blaeberries set four times more seed when crosspollinated at a 10m distance, and that cross-pollinations produced more fruit than
self-pollinations, as well (ibid., 882). Since studies like the one mentioned earlier
have determined that proximal clonal structures can show high genetic diversity,
this cannot be the only reason for smaller fruit set within short pollination
distances. Furthermore, since reproductive success and genetic diversity will
play a role in the development of Vaccinium myrtillus as a fruit crop in Scotland, a
more detailed study of this area of research is necessary, although not timepermissible for this particular work. Observation of pollination, genetics, and fruit
sets within local populations and between populations in Scotland, with regard to
the environment and other variables will be a place to start.
It is necessary to mention that Vaccinium species are known to have
mycorrhizal associations with fungi such as Pezizella ericaea and Clavaria ssp.
that infect the roots and increase the uptake of nitrogen and phosphorus from
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acidic soils (Vanderkloet 1988, 25). Vaccinium myrtillus are among the species
that share a mycorrhizal relationship, and some scholars have suggested that
this is the reason that Vaccinium species, in general, are tolerant of heavy metals
(ibid.). MacKay (2004) investigated the effects on local mycorrhizae of managed
burning on a Pinus sylvestris plantation in Inverness-shire. He found that
although he was unable to specifically identify most mycorrhizae morphotypes in
association with the pines, one, Piceirhiza bicolorata, was found associated with
the roots of both Pinus sylvestris and Vaccinium myrtillus. Previously, it was
thought that Ericoid mycorrhizae and the ectomycorhizae associated with Pinus
syvlestris were unrelated (MacKay 2004, 21). This relationship may be one
reason why blaeberries are more successful in the pine woodlands of Scotland
than on the moors. Finally, an earlier study by Turnau et. al. (1992) found that NP-K fertilization of oak and pine woodlands in southern Poland reduced the
frequency of root cells colonized with mycorrhizae. This had the effect of causing
the Vaccinium myrtillus population to disappear whilst rapidly expanding other
species like grasses Festuca gigantea and Milium effusum. Although this is a
single study from two decades ago, it underlines the importance of observing and
investigating the mycorrhizal relationship with blaeberries as part of an ongoing
research plan. The most recent issues of the International Symposium on
Vaccinium Culture (Issues 574 and 715) from the ISHS Acta Horticulturae
(http://www.ishs.org/acta/index.htm) also contain new and ongoing research into
Ericoid mycorrhizae and their relationship to Vaccinium culture.
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Appendix Three: The Flavonoid Anthocyanins
Anthocyanins are part of a much larger class of natural chemicals called
flavonoids, a vast group of aromatic plant products whose many sub-categories
include phenylpropanoids, tannins, and quinones (Brielmann 1999, 20).
Flavonoids are a product of the shikimate and acetate pathways and provide
plants with pigments that serve a variety of functions including UV ray protection,
antioxidant activity, and enzyme inhibition (Pengelly 2004, 34). The basic
flavonoid structure is two benzene rings (A and B) separated by a propane unit
(See Figure One). The three-carbon chain is often closed to form a heterocyclic
third ring (C-ring).
Figure One: Flavonoid Structure
http://www.uky.edu/~dhild/biochem/17/skeleton.gif
Flavonoids are water-soluble and mostly occur in nature as glycosides (Dewick
2001, 150). Research has shown them to be ‘good scavengers of free radicals
due to high reactivities of their hydroxyl substituents in a hydrogen atom
abstraction reaction’ (Korkina and Afanas’ev 1997, 152). Furthermore, they have
been shown to have superoxide inhibition activity through a variety of
mechanisms incuding xanthine oxidase and superoxide dismutase (ibid). They
have also been shown to inhibit prostaglandin up-regulation, inhibit cell
proliferation in tumour cell lines, and have antimutagenic properties in human
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lymphocytes (Korkhina and Afanas’ev 1997, 159). For these reasons, several
classes of flavonoids are important to human health maintenance including the
flavonols, flavones, flavonones, and anthocyanidins (Lila 2004, 306).
Since the second World War, when British RAF pilots claimed that eating
bilberry jam helped improve their night vision (Muth et al. 2000, 164), biochemists
and medical researchers have become increasingly interested in the compounds
contained in bilberries and other red, purple, and blue fruits. Much of this interest
lies in the antioxidant qualities of some of the phytochemicals bilberries contain,
namely the flavonoid anthocyanins. Anthocyanins, whose name is derived from
the Greek words for flower (anthos) and blue (kyanos), are water-soluble
pigments that provide flowers, leaves, and fruits with a wide range of colours
ranging from pink, red, and purple to blue and black. The bright colour and low
toxicity of anthocyanins plays an important role in aiding plant pollination by
insects and seed dispersal, thus reproduction, thorough foraging mammals and
birds (Bruneton 1999, 356). Anthocyanins have a basic flavonoid inner structure
of 3 carbon rings to which a number of sugar molecules may be attached,
particularly at hydroxylated positions 3, 5, and 7 on the A and heterocyclic rings
(See Figure Two).
Figure Two
http://wwwchem.uwimona.edu.jm:1104/spectra/chime/anthocy.gif
In strong acidic media (pH<3), anthocyanins are stable (and red), but they
become less stable (and blue) in weak acids (pH 4 – 6), and the maintenance of
their colour in living media may be due to a number of other mechanisms
including intermolecular co-pigmentation with other molecules like flavonoids,
74
proteins, and pectins (Bruneton 1999, 358). Anthocyanins are also unstable in
oxygen, heat, light, and in reaction with sulfur dioxide (ibid.).
The inner structure of the anthocyanin is called the anthocyanin aglycon or
anthocyanidin. Cyanidin, the most abundant anthocyandin found in nature, is
present in 69% of fruits and 50% of flowers (Evans 2002, 250). Pelargonidin and
delphinidin (See Figure Three) are also very common, ‘virtually ubiquitous’
(Bruneton 1999, 356), but peonidin, petunidin, and malvidin are also known.
Figure Three: Structures of some common anthocyanidins
http://www.shieldsgardens.com/DLPlace/anthocyanidins.jpg
The anthocyanidins themselves are unstable because their 3-hydroxyl group
makes them very reactive, therefore they are never found in nature
unglycosylated (Bruneton 1999, 357). Anthocyandins may be attached to a
number of different sugars, including glucose, galactose, rhamnose, and
arabinose, and are often known by the number of sugar molecules in their
structure (e.g. monosides, biosides, triosides) (Lala 2005, 16). Their diversity
can be attributed to the varying number of sugar molecules in their structure and
by chemical combination of these sugars with organic acids like phenylpropanoic
or benzoic acid (ibid., 14). It is important to take notice of the differences
between anthocyanins and anthocyanidins and when they occur because this is
75
a matter of considerable importance when considering their pharmacological
applications. Similarly, it should be noted that anthocyanin solutions are very
unstable, and can only be kept in nitrogen, at low temperature and in the dark
(Bruneton 1999, 359.). This has obvious ramifications for both their experimental
usage and their pharmacological and food industry applications.
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