Traditional food:
functional properties and perspectives
Diana Di Gioia
Department of Agroenvironmental Science and Technology
Alma Mater Studiorum-University of Bologna - Italy
1
The Agriculture Faculty of UNIBO
Department of Agroenvironmental Science and Technology
D. Di Gioia, B. Biavati, G. Dinelli, I. Marotti,
Microbiology
Agronomycrop physiology
2
What is a “traditional food”?
Different countries have differences in the definition and
characterization of Traditional Food Products (TFPs)
depending on cultural heritage, regional specificities and
many other factors
For the European consumers:
Food produced,
processed
and can
prepared
The geographical
link often
“food products that are well-known,
that
one
eat
in a must
given geographical
least
It doesby
notour
refergrandparents
to theoccur in at ”.
and that were eaten already
areaone
using
recognised
stage
of the
origin but highlights
know-how
production
traditional character
The European legal framework:
either in the composition
means
of production
Protected Designationorof
Origin
(PDO), along
with
Protected Geographical Indication (PGI) and Traditional
Speciality Guaranteed (TSG), are systems created by the
European Union in 1992 to promote and protect regional food
products within the EU.
3
Unique definition for TFPs?
traditional
vs
Time immutable?
modern
No history?
Hamburger
Mongols (1200)
Tartare stick
Mortadella: sausage (PGI)
("The
Ricottacalled
eaters")
Vincenzo
Campi,
~1585
The
Romans
the
sausage
"farcimen
mirtatum" (myrtle sausage), because the
sausage was flavored with myrtle berries.
Immigrants (1850)
W
i
m
p
y
(1930)
4
Content
The aim is to give an overview of the functional
properties of TFPs
 The health image of TFPs: the role of plant
ingredients
 The health image of TFPs: the role of
microorganisms
The role of TFPs against obesity
 Challenges and perspectives of the TFP sector
5
 The health image of TPFs: the
role of plant ingredients
Recent surveys have evidenced that EU consumers
associate several attributes to TFPs : are the presumed
“health benefits” of TFPs confirmed by scientific data?
Traditional ingredients (for example old vs modern
crop genotypes)
Traditional production of ingredients (for example
organic vs conventional cropping systems)
6
The health image of TPFs: the role of plant ingredients
Comparison of functional properties (bioactive compounds)
of old and modern varieties of common and durum wheat
Old varieties
Agronomic drawbacks (not dwarf,
lodging, weed resistance)
Adapted to “low input” farming
Old cultivar
Modern cultivar
Modern varieties
Introduced after the “green revolution” (1955-1970)
Extraordinary yield and optimal rheological properties
Adapted to “high input” farming (fertilizers, herbicides)
Relatively low genetic variability
7
The health image of TPFs: the role of plant ingredients
Lignans are important bioactive compounds in wheat
Lignans pharmacological properties:
 anti-inflammatory activity
 immunosuppressive activity
 anti-oxidant activity
 antiviral actions
 protective effect against estrogen-related
disease such as osteoporosis
 anti-cancer properties (reduced incidence
rate of breast, prostate and colon cancer)
8
The health image of TPFs: the role of plant ingredients
Lignan content (µg/g dry weight) in 10 soft wheat varieties
ARC
HIN
SYR
SECO
(tot1)
PIN+ISO
Total
Bolero
n.d.
n.d.
n.d.
1.55 ± 0.13
1.22 ± 0.10
2.77 ± 0.16
Mieti
n.d.
n.d.
n.d.
1.64 ± 0.08
1.02 ± 0.08
2.66 ± 0.12
Nobel
n.d.
n.d.
n.d.
1.30 ± 0.15
0.98 ± 0.08
2.29 ± 0.16
Eureka
n.d.
n.d.
n.d.
1.58 ± 0.15
1.09 ± 0.07
2.67 ± 0.04
Verna
1.02 ± 0.11
0.95 ± 0.10
n.d.
1.44 ± 0.12
1.22 ± 0.07
4.64 ± 0.14
Sieve
n.d.
1.00 ± 0.08
n.d.
1.34 ± 0.11
1.01 ± 0.10
3.36 ± 0.04
Andriolo
0.96 ± 0.12
1.00 ± 0.10
n.d.
1.33 ± 0.08
1.09 ± 0.08
4.38 ± 0.10
Inallettabile
0.85 ± 0.08
0.84 ± 0.06
1.34 ± 0.11
1.53 ± 0.12
1.28 ± 0.07
5.84 ± 0.11
Gentil Rosso
aristato
Gentil Rosso
mutico
1.08 ± 0.08
0.93 ± 0.07
n.d.
1.25 ± 0.13
1.42 ± 0.10
4.67 ± 0.11
1.20 ± 0.08
1.59 ± 0.10
1.35 ± 0.14
2.46 ± 0.14
1.86 ± 0.09
7.11 ± 0.05
(tot1)= sum of SECO and ANYDROSECO
Mean
content:
2.60 ± 0.21
Mean
content:
5.00 ± 1.30
Dinelli et al. (2007), Electrophoresis, 28, 4212-4219. 9
The health image of TPFs: the role of plant ingredients
Free polyphenols
(mg/100g)
Bound polyphenols
(mg/100g)
Total polyphenols
(mg/100g)
Modern cv. (6)
45.5 ± 3.0
178.0 ± 12.4
223.5 ± 12.1
Old cv (17)
60.5 ± 3.5
263.1 ± 15.7
313.6 ± 15.9
Free flavonoids
(mmol/g)
Bound flavonoids
(mmol/g)
Total flavonoids
(mmol/g)
Modern cv. (6)
0.41 ± 0.12
0.84 ± 0.20
1.25 ± 0.21
Old cv (17)
0.51 ± 0.13
1.41 ± 0.25
1.92 ± 0.22
Total carotenoids
(mg/100g)
Lutein
(mg/100g)
Resistant starch
(%)
Modern cv. (6)
149.9 ± 26.5
70.0 ± 8.5
2.83 ± 0.49
Old cv (17)
232.9 ± 14.0
91.9 ± 9.5
4.09 ± 0.83
Dinelli et al. (2009), submitted to Food chem
10
The health image of TPFs: the role of plant ingredients
General considerations on old crop genotypes:
They did not undergo strict breeding programs and thus they
are characterized by a wide genetic variability;
Modern crop breeding programs are often aimed to reduce the
crop carbon investment in secondary metabolites (=functional
compounds);
Old genotypes are an important biodiversity source to be used
in organic farming and to increase the content of healthpromoting compounds.
Plant breeding: the art and science of changing the genetics of plants for
the benefit of humankind (Poehlman & Sleeper, Breeding Field Crops, 1995)
11
The health image of TPFs: the role of plant ingredients
Wounding
UV Light
anthocyanidins
flavonols/ flavones
cholorogenic acid
coumarins
lignin
psoralens
Pathogen Attack
isoflavones
Low Nitrogen
coumarins
flavonoids
flavonols
isoflavonoids
stilbenes
It is recognized that highly-intensive agricultural practices (conventional
farming) may reduce
metabolites
involved
Lowthe
Ironnatural production of secondary
Low
Phosphate
in plant defense;
phenolic acids
anthocyanidins
Under organic farming conditions, plants are easily subject to stress
conditions  elicitation effects on secondary metabolites  increment in
Dixon & Paiva, The plant cell, 1995, 7; 1085-1097.
functional compounds.
12

The health image of TPFs: the role of
microorganisms
Beneficial bacteria in the food chain can have:
BIOPRESERVATION:
life and
a protective role (protectiveexhended
cultures),storage
to reduce
the
safety of foodsin
growth of pathogenic and/orenhanced
spoilage microrganisms
foodBIOPRESERVATION using their natural or
controlled microflora and
(or) their antibacterial
a probiotic role (probiotic cultures),
products i.e. conferring a
beneficial effect upon the host, either on a farm animal
through animal feed, or on humans, through different
food products.
13
The health image of TFPs: the role of microorganisms
LAB and bifidobacteria are the best candidates for use as protective
& probiotic cultures:
 Have been used since the beginning of history as starter cultures
 Present in almost all fermented foods-vegetables, meat products,
dairy products
 Are part of the natural microbiota of both farm animals and humans
 Have a long history of consumption and safe use
Lactobacillus
Bifidobacterium
Pediococcus
Lactococcus
Streptococcus
Oenococcus
14
The health image of TFPs: the role of microorganisms
Relevance of LAB in different fermented food products
Wine
Beer
Cider
Cheese
Sauerkraut
Fermented
milk
Fermented
sausages
15
The health image of TFPs: food preparation
Protective cultures: applications in traditional dairy products
Soft cheeses
 Control of pathogens such as Listeria spp.
Mascarpone
Brie
Soft Cheese
Semi hard & hard cheeses
 Growth control of undesirable spoilage bacteria
Cheddar
Parmiggiano
Reggiano
Yogurt
The starter cultures used
can be considered protective
cultures
Starter cultures can also
have a role as probiotic
cultures
Pecorino
16
The health image of TFPs: the role of microorganisms
Protective cultures: applications in traditional meat products
Cured meat, dry or semi dry
 Control of pathogens such as Campylobacter spp.
Beef jerky
Bresaola
Semi dry sausages
Cooked or ground meat
 Control of pathogens such as Campylobacter spp.
Cured meats
Ground meat
Poultry
17
The health image of TFPs: the role of microorganisms
Protective/probiotic cultures:
applications in traditional meat products
 Campylobacteriosis is the most frequently reported zoonotic
disease in Europe
 the use of antibiotics in feed to prevent colonization of
Campylobacter spp. has been prohibited in Western Europe
 alternative strategies to contrast the infection from this
bacterium in animals have to be identified and used
 there is an increasing interest in the development of alternative
therapies based on the use of probiotics derived from the natural
intestinal flora to contrast the infection of this bacterium
“Control and Prevention of Emerging and Future Pathogens at
Cellular and Molecular Level throughout the Food Chain”
18
The health image of TFPs: the role of microorganisms
Study of the role of protective and probiotic cultures in food
MAIN OBJECTIVES OF THE WORK:
To select protective and probiotic cultures of LAB and bifidobacteria able to:
- inhibit pathogenic organisms common in the food industry (Campylobacter
jejuni)
- survive food processing conditions (presence of salt, high temperature, lack
of nutrients)
- survive the passage in the gastrointestinal tract of animals or humans (low
pH, presence of bile salts)
To evaluate the capacity of orally administered probiotics (selected as active
against Campylobacter jejuni ) to colonise the intestinal tract of broiler chickens
and assess their effect on the Campylobacter jejuni population
19
The health image of TFPs: the role of microorganisms
Selection of protective and probiotic cultures of LAB and
bifidobacteria able to inhibit Campylobacter jejuni strains
92 strains
(LAB and bifidobacteria)
were tested against:
C. jejuni CIP 70.2
C jejuni LMG 8842
C. jejuni 221/05
20
The health image of TFPs: the role of microorganisms
Screening methods:
 agar spot assay with LAB and bifidobacteria cultures
 well diffusion agar assay using neutralized cell-free cultures
supernatants of the strains showing inhibitory activity
Antimicrobial activity against
Spot of culture
Campylobacter
spp.
Nutrient soft agar (0,5 %)
containing about 107 cells per ml
of C. jejuni
Lactobacillus delbrueckii
ssp delbrueckii PCK 103
L. plantarum
PCK 73
B. Longum
PCB 133
21
The health image of TFPs: the role of microorganisms
Inhibitory activity: well diffusion agar assay with
neutralized cell-free surnatants
Campylobacter
jejuni CIP 70.1
Campylobacter
jejuni LMG 8842
Campylobacter
jejuni 221/05
L. pentosus PCA 227
-
+
-
L. plantarum PCA 236
-
+
-
L. plantarum PCA 263
-
+
+
L. plantarum PCA 275
-
+
+
L. plantarum PCS 20
-
+
+
B. longum PCD 733B
-
-
+
Enterococcus durans
PCD 103
-
+
-
Leuconostoc
mesenteroides PCK 18
-
-
+
L. plantarum PCK 73
+
-
+
L. delbrueckii PCK 103
-
+
+
B. longum PCB 133
-
+
+
22
The health image of TFPs: the role of microorganisms
Survival to food processing conditions:
6% NaCl, thermal stress (50°C and 55°C), starvation stress
• Heat Stress
all strains survive well after 1h of treatment to 55 °C
• Osmotic stress
all strains retain their viability after 1h in presence of 6% NaCl
• Starvation stress
three strains after 6 hours loose more than 50% of their initial
population. The others strains, after 24 hours, didn’t show a
significant vitality decrease
23
The health image of TFPs: the role of microorganisms
Survival the passage in the GIT of animals or humans
pH 2.5, bile salts 1 and 2%
• pH 2,5
ResistanceResistance
to 2%
bile salts
to bile salts 2%
all strains rapidly loose their
viability after 1 h of treatment
100,00
90,00
80,00
Survival %
70,00
60,00
50,00
• Bile salts 1% and 2% w/v
more than 50% of the initial
population survive after 4 hours of
treatment
40,00
30,00
20,00
10,00
0,00
0'
Minutes
60'
240'
PCA
227
PCA
236
PCA
263
PCA
275
PCK
73
PCK
18
PCK
103
PCD
733B
PCD
103
PCB
133
PCS
20
PCA 227
PCA 236
PCA 263
PCA 275
PCK 73
PCK 18
PCK 103
PCD 733B
PCD 103
PCB 133
PCS 20
Survival
%
Strains
LAB and Bifidobacteria strains
Microencapsulation of
freeze-dried cells to
improve survival
24
The health image of TFPs: the role of microorganisms
Administration in poultry of probiotic cultures selected as active
against Campylobacter jejuni
• Probiotic
administration
Bifidobacterium longum
(frozen cultures 108-109 by
oral gavage)
Lactobacillus plantarum
PCB133
PCS20
In vivo conditions for all the trials
• 15 chickens each group
• Chickens hygienic condition
• 5 days naturalization
• 10 chickens chosen for faeces
collection
The health image of TFPs: the role of microorganisms
Analysis Workflow
Daily probiotic administration
for two weeks
Samples collection: T0-T1 (15
days)-T2 (20 days)
Culture-dependent
techniques
to assess
microorganisms
vitality in
faeces
Microbiota analysis
Cultureindependent
technique
DNA extraction
Molecular analysis
Real Time-PCR
The health image of TFPs: the role of microorganisms
4
PCB133
End of administration
2
0
0
10
20
T0
T1
T2
6
Bifidobacterium longum (n. cell/g faeces)
Mean values (log cell/g of faeces)
B. longum colonization
4
2
0
N.22
N.11
N.21
N.30
N.4
N.41
N.47
N.45
N.18
N.17
N. chickens
Time (Days)
Only the probiotic strain B. longum PCB133 colonised the intestinal tract
of the broiler chickens and was detected in the faeces of the treatment
group
L. Plantarum PCS20 did not
Sensible reduction of C. jejuni
(preliminary results to be confirmed)
Culture-dependent vs culture-indipendent technique ~ 1 log difference
The health image of TFPs: the role of microorganisms
Work in progress.....
 Impact of two prebiotic additives on the
intestinal microbiota of poultry
 Detection of pathogens in animal feed by
RealTime PCR and evaluation of the survival of the
microencapsulated probiotics
 Application of a new synbiotic formula in the
feeding of poultry with relevance to microbiological
aspects and growth performance
28
The health image of TFPs: the role of microorganisms
Interaction between probiotics and plant derived
functional componds
Flavonoids are a large
group of plant
secondary metabolites,
which belong to the
class/category of
polyphenols. They are
omnipresent in the
human diet and are
found in fruits,
vegetables and grains.
Flavonoids in food are generally
considered to be beneficial to
consumers' health and are one of the
most important categories of bioactive
food components.
Epidemiological observations ascribed
to flavonoids:
to boost the immune sytem
to protect body from free radicals
To kill pathogenic germs
To decrease the risk of cancer
29
The health image of TFPs: the role of microorganisms
Flavonoid bioconversion by microorganisms
Dietary
glycosides
Aglycones
Bacterial
hydrolysis
Bioactive form
Glucosidase
Fla Glu
Fla
Absorption
Glu
Bacterial hydrolysis
Phenolic acid
CO2
Absorption
The health image of TFPs: the role of microorganisms
Flavonoid bioconversion by the gut microflora
Bioavailability of dietary flavonoids 
ability of gut microflora to degrade these
compounds (b-glucosidase activity)
Several groups of bacteria
possess b-glucosidase activity
(Bacteroides, Lactobacillus,
Bifidobacterium)
Gut
microflora
Bifidobacterium Lactobacillus
Widely used in pharmaceutical and dairy preparations for their
probiotic effects (maintain improved intestinal bacteria composition,
stimulate the immune response, possible anticarcinogenic activity
and protection against infections).
The health image of TFPs: the role of microorganisms
Flavonoid bioconversion by the gut microflora
Aim of the work
1) Screening of Bifidobacterium strains
for b-glucosidase activity
2) Capability of Bifidobacterium strains to convert the
flavonoid glycosides detected in common bean seeds and
seedlings
• 38 bifidobacteria strains were
screened for b-glucosidase
activity
•B. catenulatum
•B. pseudocatenulatum
•B. adolescentis
•B. longum
•B. infantis
•B. breve
Flavonoid glycosides
• Authentic standards
(kaempferol 3-O-glucoside,
kaempferol 3-Oxylosylglucoside, daidzin,
glycitin, genistin) of the Italian
common bean ecotype
“Zolfino”
. Seed and seedling extracts
The health image of TFPs: the role of microorganisms
Screening of b-glucosidase activity
determination of p-nitrophenyl β-D-glucopiranoside conversion
in p-nitrophenol.
Selected bacteria
(pure culture)
Flavonoid glycosides
Standards
Bean seeds
Incubation (37°C, anerobiosis, 24h)
Bacterial surnatants
Flavonoid extraction (SPE)
HPLC analysis
40.0
30.0
20.0
B7377
B7304
B7003
B7178
B7291
B7184
B7001
B7875
B7039
B7824
B7239
B7370
B7023
B7696
B7792
B7031
B7716
B7819
B7740
B7162
B7284
B7305
B7314
B7290
B7310
B7308
B7296
B7229
B7213
B7262
B7249
B7825
B7352
B7348
B7302
B7254
B7356
B7751
β-glucosidase activity (U/g dry cell weight)
The health image of TFPs: the role of microorganisms
Screening of b-glucosidase activity: results
50.0
= B. catenulatum
= B. adolescentis
= B. pseudocatenulatum
= B. infantis
= B. breve
= B. bifidum
= B. longum
10.0
0.0
*
*
30.0
*
*
*
50.0
B7377
B7304
B7003
B7178
B7291
B7184
B7001
B7875
B7039
B7824
B7239
B7370
B7023
B7696
B7792
B7031
B7716
B7819
B7740
B7162
B7284
B7305
B7314
B7290
B7310
B7308
B7296
B7229
B7213
B7262
B7249
B7825
B7352
B7348
B7302
B7254
B7356
B7751
β-glucosidase activity (U/g dry cell weight)
The health image of TFPs: the role of microorganisms
b-glucosidase activity: results
B. catenulatum
B. adolescentis
40.0
B. pseudocatenulatum
20.0
B. infantis
B. breve
10.0
0.0
The health image of TFPs: the role of microorganisms
Flavonoid metabolism: selected bifidobacteria strains incubated with
flavonoids extracted from “Zolfino” bean seeds
Kaempferol
kaempferol 3-O-xylosylglucoside
kaempferol 3-O-glucoside
Kaempferol-3-0-xylosylglucoside
kaempferol 7-O-glucoside
AFTER incubation with B7003
0
4
Time
8
12
The health image of TFPs: the role of microorganisms
Flavonoid metabolism: selected bifidobacteria strains
incubated with flavonoids extracted from “Zolfino” bean
seedlings
Glycosidic forms of daidzein,
genistein, glycitein,
kaempferol
Genistein
Glycitein
Chromatogram
after 24 h incubation
with B7003
Daidzein
Chromatogram
Seedling extract
Zolfino landrace
0
Kaempferol
4
Time (min)
8
12
The health image of TFPs: the role of microorganisms
Degradation rate k (1/h)
Metabolism of flavonoid aglycones  degradation rates following
incubation with selected strains
0.25
0.20
0.15
Daidzein
Genistein
Glycitein
Kaempferol
0.10
0.05
0.00
Daidzein
Genistein
Glycitein
Kaempferol
Correlation between chemical structure and rate of microbial
degradation: number and position of hydroxyl groups are important
structural characteristics for flavonoid degradation by human bifidobacteria:
genistein and kaempferol were degraded faster than daidzein and glycitein.
Marotti I. et al. J. Agric. Food Chem. 2007, 55, 3913-3919
The health image of TFPs: the role of microorganisms
= L. plantarum
10
= L. kefiri
8
= L. buchneri
= L. lactis
6
= L. acidophilus
4
= L. helveticus
= L. fermentum
2
= L. bulgaricus
L
L
L
L
L
L
L
L
L
L
3
8
L
2
7 12
L
10 15 9 13 11
L
5
0
14 1
β-glucosidase activity (U/g dry cell weight)
Screening of β-glucosidase activity in Lactobacillus spp.:
preliminary results
Studies in progress: wider screening of Lactobacillus strains
The health image of TFPs: the role of microorganisms
PERSPECTIVES
Mediterranean diet: bean-based dishes in the traditional
cooking common bean is a valuable ingredient to obtain
products with health benefits
Selected Bifidobacterium strains  probiotic dietary
adjuncts to improve the organoleptic properties and health
beneficial effects of flavonoid-based products, including
hypothetical common bean food derivatives
Potentiality of investigated Lactobacillus strains as functional
starter cultures for new fermented products  facilitate
bioavailability
Roles of TFPs against “obesity”
Obesity is associated
• dietary with
abundance
significant risks of ill health
• sedentary lifestyle
41
0
Average
1167
Prentice & Jebb (2003), Obesity reviews, 4, 187-194.
1054
439
Meals
Staples
Stauces
Breakfast
1087
Muffin/Danish
Fries (chips)
Chicken McNuggets
Dressings
Breakfast
Sauces
Dessert/Shakes
Sandwiches
Salads
Desserts
Fries (chips)
Sandwiches
Fried + side
Condiments, etc
Sandwiches
Breakfast
Popcorn
Wings
Biscuit
Desserts
Chicken
Fries (chips)
Crispy strips
Sandwiches
Vegetables
Energy density (kJ 100 g-1)
Roles of TFPs against “obesity”
Overnutrition and energy density
Outlet A
Outlet B
Outlet C
Burger King
Jack in the Box
Mc Donald’s
Traditional
Gambian
1600
1200
800
British
diet (670)
400
42
Roles of TFPs against “obesity”
Overnutrition and energy density
5000
For assuring 8500 kJ per day
Traditional
Gambian
Weight of food eaten (g)
4000
3000
If a person made a 200 g error on a
diet of 1200 kJ 100 g-1 when eating
fast foods just twice a week, the
cumulative effect would add up to
250000 kJ per year – equivalent to
almost 8 kg of adipose tissue.
Market
(ready dishes)
2000
Fast foods
1000
0
0
400
800
1200
1600
-1
Energy density (kJ 100 g )
Prentice & Jebb (2003), Obesity reviews, 4, 187-194.
2000
43
Roles of TFPs against “obesity”
Obesity predisposes the individual to the development of nontransmissible diseases
The role of traditional Mediterranean diet
Whole cohort set
Mediterranean diet
• high intake
of vegetables, legumes, fruit, and cereals
Greece
(in the past
Spain largely unrefined);
• moderate
Italy to high intake of fish;
• low intake
France of saturated lipids but high intake of
unsaturated
UK lipids (olive oil)
Denmark intake of dairy products (mostly cheese and
• moderate
Germany
yogurt)
Sweden of meat
• low intake
Netherlands
0
1
2
3
4
5
6
7
Mortality ratios (%)
Trichopoulou et al., (2005), BMJ, 330, 991-997.
44
Perspectives and challenges of TFP sector
A traditional food is not “innovative” but ……
…..innovations are costantly looked for to
 guarantee the safety of the products (ensuring
safety through innovation)
 improve the nutritional and health properties of the
food (link between nutrition and health)
 preserve biodiversity and sustainability
….. this is what the modern consumer and the modern
scientist always keep in mind……
45
Perspectives and challenges of TFP sector
Value (Technology +Iinformation)
Personalized Nutrigenetic Food Age
Nutrigenetic
food
Functional
food
Convenience
food
Molecular nutrition
Tailor-made personal food
Home-test kits
Chemical analysis
Nutraceutical
Target group
Health claim
Essential nutrients
Engineered food
Mass production
Survival food
Future
18th
19th
20th
21th Century
46
Perspectives and challenges of TFP sector
Personalized Nutrigenetic Food Age
TFPs
Pathologic status
Tailor made diet
Health status
Comparison
47
Thank you
48