traditional food: functional properties and perspectives diana di gioia 1 department of...
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Traditional food:functional properties and
perspectives
Diana Di Gioia
1
Department of Agroenvironmental Science and Technology
Alma Mater Studiorum-University of Bologna - Italy
2
The Agriculture Faculty of UNIBO
Microbiology Agronomy-crop physiology
D. Di Gioia, B. Biavati, G. Dinelli, I. Marotti,
Department of Agroenvironmental Science and Technology
What is a “traditional food”?
3
For the European consumers: “food products that are well-known, that one can eat often and that were eaten already by our grandparents ”.The European legal framework: Protected Designation of 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.
Different countries have differences in the definition and characterization of Traditional Food Products (TFPs) depending on cultural heritage, regional specificities and many other factors
Food produced, processed and preparedin a given geographical area using recognised know-how
The geographical linkmust occur in at least one stage of the production
It does not refer to the origin but highlights traditional character either in the compositionor means of production
Unique definition for TFPs?
4
("The Ricotta eaters") Vincenzo Campi, ~1585
traditional vs modern
Time immutable?
Mortadella: sausage (PGI)
The Romans called the sausage "farcimen mirtatum" (myrtle sausage), because the sausage was flavored with myrtle berries.
No history?Hamburger
Mongols (1200)
Tartare stick
Wimpy
(1930)
Immigrants (1850)
Content
5
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
Challenges and perspectives of the TFP sector
The role of TFPs against obesity
The health image of TPFs: the role of plant ingredients
6
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)
7
Comparison of functional properties (bioactive compounds) of old and modern varieties of common and durum wheat
Modern cultivar
Introduced after the “green revolution” (1955-1970)Extraordinary yield and optimal rheological properties
Adapted to “high input” farming (fertilizers, herbicides)Relatively low genetic variability
Modern varieties
Agronomic drawbacks (not dwarf, lodging, weed resistance)
Adapted to “low input” farming Old cultivar
Old varieties
The health image of TPFs: the role of plant ingredients
8
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)
The health image of TPFs: the role of plant ingredients
9
Lignan content (µg/g dry weight) in 10 soft wheat varieties
Mean content:
2.60 ± 0.21
Mean content:
5.00 ± 1.30
(tot1)= sum of SECO and ANYDROSECO Dinelli et al. (2007), Electrophoresis, 28, 4212-
4219.
2.67 ± 0.041.09 ± 0.071.58 ± 0.15n.d.n.d.n.d.Eureka
2.29 ± 0.16
0.98 ± 0.081.30 ± 0.15n.d.n.d.n.d.Nobel
2.66 ± 0.121.02 ± 0.081.64 ± 0.08n.d.n.d.n.d.Mieti
2.77 ± 0.161.22 ± 0.101.55 ± 0.13n.d.n.d.n.d.Bolero
TotalPIN+ISOSECO (tot1)
SYRHINARC
5.84 ± 0.111.28 ± 0.071.53 ± 0.121.34 ± 0.110.84 ± 0.060.85 ± 0.08Inallettabile
4.38 ± 0.101.09 ± 0.081.33 ± 0.08n.d.1.00 ± 0.100.96 ± 0.12Andriolo
3.36 ± 0.041.01 ± 0.101.34 ± 0.11n.d.1.00 ± 0.08n.d.Sieve
4.64 ± 0.141.22 ± 0.071.44 ± 0.12n.d.0.95 ± 0.101.02 ± 0.11Verna
7.11 ± 0.051.86 ± 0.092.46 ± 0.141.35 ± 0.141.59 ± 0.101.20 ± 0.08
4.67 ± 0.111.42 ± 0.101.25 ± 0.13n.d.0.93 ± 0.071.08 ± 0.08Gentil RossoaristatoGentil Rossomutico
The health image of TPFs: the role of plant ingredients
10
45.5 ± 3.0Modern cv. (6)
60.5 ± 3.5 313.6 ± 15.9Old cv (17)
Free polyphenols(mg/100g)
Bound polyphenols(mg/100g)
Total polyphenols(mg/100g)
263.1 ± 15.7
178.0 ± 12.4 223.5 ± 12.1
0.41 ± 0.12Modern cv. (6)
0.51 ± 0.13 1.92 ± 0.22Old cv (17)
Free flavonoids(mol/g)
Bound flavonoids(mol/g)
Total flavonoidsmol/g)
1.41 ± 0.25
0.84 ± 0.20 1.25 ± 0.21
Modern cv. (6)
Old cv (17)
149.9 ± 26.5
232.9 ± 14.0
Total carotenoids (g/100g)
4.09 ± 0.83
Resistant starch(%)
2.83 ± 0.49
Lutein(g/100g)
91.9 ± 9.5
70.0 ± 8.5
Dinelli et al. (2009), submitted to Food chem
The health image of TPFs: the role of plant ingredients
11
They did not undergo strict breeding programs and thus they are characterized by a wide genetic variability;
General considerations on old crop genotypes:
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 health-promoting compounds.
Plant breeding: the art and science of changing the genetics of plants for the benefit of humankind (Poehlman & Sleeper, Breeding Field Crops, 1995)
The health image of TPFs: the role of plant ingredients
12
Wounding
cholorogenic acid
coumarins
lignin
Dixon & Paiva, The plant cell, 1995, 7; 1085-1097.
Low Nitrogen
flavonoids
isoflavonoids
UV Light
anthocyanidins
flavonols/ flavones
psoralens
Pathogen Attack
isoflavones
coumarins
flavonols
stilbenes
Low Phosphate
anthocyanidins
Low Iron
phenolic acids
It is recognized that highly-intensive agricultural practices (conventional farming) may reduce the natural production of secondary metabolites involved in plant defense;
Under organic farming conditions, plants are easily subject to stress conditions elicitation effects on secondary metabolites increment in functional compounds.
13
Beneficial bacteria in the food chain can have:
a protective role (protective cultures), to reduce the growth of pathogenic and/or spoilage microrganisms in foodBIOPRESERVATION
a probiotic role (probiotic cultures), i.e. conferring a beneficial effect upon the host, either on a farm animal through animal feed, or on humans, through different food products.
The health image of TPFs: the role of microorganisms
BIOPRESERVATION: exhended storage life and enhanced safety of foods using their natural or controlled microflora and (or) their antibacterial products
14
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
Lactococcus
Streptococcu
s
Pediococcu
s
Oenococcu
s
Bifidobacterium
The health image of TFPs: the role of microorganisms
15
Relevance of LAB in different fermented food products
Wine BeerCider
Sauerkraut
Fermented milk
Cheese Fermentedsausages
The health image of TFPs: the role of microorganisms
16
The health image of TFPs: food preparation
Protective cultures: applications in traditional dairy products
YogurtThe starter cultures used can be considered protective culturesStarter cultures can also have a role as probiotic cultures
BrieMascarpone Soft Cheese
Soft cheeses Control of pathogens such as Listeria spp.
Cheddar Pecorino
Semi hard & hard cheeses Growth control of undesirable spoilage bacteria
Parmiggiano Reggiano
17
Protective cultures: applications in traditional meat products
Beef jerky Bresaola Semi dry sausages
Cured meat, dry or semi dry Control of pathogens such as Campylobacter spp.
Cooked or ground meat Control of pathogens such as
Campylobacter spp.
Cured meats Ground meat Poultry
The health image of TFPs: the role of microorganisms
18
The health image of TFPs: the role of microorganisms
Protective/probiotic cultures: applications in traditional meat products
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”
Campylobacteriosis is the most frequently reported zoonotic disease in Europe
19
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)
Study of the role of protective and probiotic cultures in food
The health image of TFPs: the role of microorganisms
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
20
92 strains (LAB and bifidobacteria)
were tested against:
C. jejuni CIP 70.2 C jejuni LMG 8842
C. jejuni 221/05
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
21
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
Nutrient soft agar (0,5 %) containing about 107 cells per ml
of C. jejuni
Spot of culture
The health image of TFPs: the role of microorganisms
Lactobacillus delbrueckii ssp delbrueckii PCK 103
L. plantarum PCK 73
B. Longum PCB 133
Antimicrobial activity against Campylobacter spp.
22
Inhibitory activity: well diffusion agar assay with neutralized cell-free surnatants
The health image of TFPs: the role of microorganisms
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 - + +
23
Survival to food processing conditions:
6% NaCl, thermal stress (50°C and 55°C), starvation stress
The health image of TFPs: the role of microorganisms
• 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
24
Survival the passage in the GIT of animals or humans
pH 2.5, bile salts 1 and 2%
The health image of TFPs: the role of microorganisms
• pH 2,5 all strains rapidly loose their viability after 1 h of treatment
• Bile salts 1% and 2% w/v more than 50% of the initial population survive after 4 hours of treatment 0'
60'240'
PCA227
PCA236
PCA263
PCA275
PCK73
PCK18
PCK103
PCD733B
PCD103
PCB133
PCS20
0,00
10,00
20,00
30,00
40,00
50,00
60,00
70,00
80,00
90,00
100,00
Minutes StrainsLAB and Bifidobacteria strains
Resistance to 2% bile salts
Survival %
Microencapsulation of freeze-dried cells to improve survival
Administration in poultry of probiotic cultures selected as active against Campylobacter jejuni
• Probiotic administration(frozen cultures 108-109
by oral gavage)
Bifidobacterium longum
PCB133
Lactobacillus plantarum
PCS20
• Chickens hygienic condition
• 5 days naturalization
In vivo conditions for all the trials• 15 chickens each group
• 10 chickens chosen for faeces collection
The health image of TFPs: the role of microorganisms
Molecular analysis
Daily probiotic administration for two weeks
Samples collection: T0-T1 (15 days)-T2 (20 days)
DNA extraction
Microbiota analysisCulture-dependent
techniques
Culture-independent
technique
Analysis Workflow
to assess microorganisms vitality in
faeces
The health image of TFPs: the role of microorganisms
Real Time-PCR
B. longum colonization
L. Plantarum PCS20 did not
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
End of administration
Culture-dependent vs culture-indipendent technique ~ 1 log difference
Sensible reduction of C. jejuni (preliminary results to be confirmed)
The health image of TFPs: the role of microorganisms
28
The health image of TFPs: the role of microorganisms
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
Work in progress.....
29
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. to boost the immune sytem
to protect body from free radicals
To kill pathogenic germs
To decrease the risk of cancer
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:
Dietary glycosides
Fla Glu
CO2 Phenolic acid
Bacterial hydrolysis
Aglycones Bacterial hydrolysis
Glu
FlaGlucosidase
Absorption
Bioactive form
Absorption
Flavonoid bioconversion by microorganisms
The health image of TFPs: the role of microorganisms
Gut Gut microfloramicroflora
Bioavailability of dietary flavonoids ability of gut microflora to degrade these compounds
glucosidase activity)
Several groups of bacteria possess -glucosidase activity (Bacteroides, Lactobacillus, Bifidobacterium)
The health image of TFPs: the role of microorganisms
Bifidobacterium
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).
Lactobacillus
Flavonoid bioconversion by the gut microflora
Flavonoid glycosides• Authentic standards (kaempferol 3-O-glucoside, kaempferol 3-O-xylosylglucoside, daidzin, glycitin, genistin) of the Italian common bean ecotype“Zolfino”. Seed and seedling extracts
• 38 bifidobacteria strains were screened for -glucosidase activity•B. catenulatum•B. pseudocatenulatum•B. adolescentis•B. longum•B. infantis•B. breve
1) Screening of Bifidobacterium strains for -glucosidase activity
2) Capability of Bifidobacterium strains to convert the flavonoid glycosides detected in common bean seeds and seedlings
The health image of TFPs: the role of microorganisms
Aim of the work
Flavonoid bioconversion by the gut microflora
Bacterial surnatants
Selected bacteria(pure culture)
Flavonoid glycosides
Standards Bean seeds
Incubation (37°C, anerobiosis, 24h)
Flavonoid extraction (SPE)
HPLC analysis
Screening of -glucosidase activity determination of p-nitrophenyl β-D-glucopiranoside conversion in p-nitrophenol.
The health image of TFPs: the role of microorganisms
0.0
10.0
20.0
30.0
40.0
50.0
B7
37
7B
73
04
B7
00
3B
71
78
B7
29
1B
71
84
B7
00
1B
78
75
B7
03
9B
78
24
B7
23
9B
73
70
B7
02
3B
76
96
B7
79
2B
70
31
B7
71
6B
78
19
B7
74
0B
71
62
B7
28
4B
73
05
B7
31
4B
72
90
B7
31
0B
73
08
B7
29
6B
72
29
B7
21
3B
72
62
B7
24
9B
78
25
B7
35
2B
73
48
B7
30
2B
72
54
B7
35
6B
77
51
β-g
luco
sid
ase
acti
vity
(U
/g d
ry c
ell
wei
gh
t)
= B. catenulatum
= B. adolescentis
= B. pseudocatenulatum
= B. infantis
= B. breve
= B. bifidum
= B. longum
The health image of TFPs: the role of microorganisms
Screening of -glucosidase activity: results
0.0
10.0
20.0
30.0
40.0
50.0
B7
37
7B
73
04
B7
00
3B
71
78
B7
29
1B
71
84
B7
00
1B
78
75
B7
03
9B
78
24
B7
23
9B
73
70
B7
02
3B
76
96
B7
79
2B
70
31
B7
71
6B
78
19
B7
74
0B
71
62
B7
28
4B
73
05
B7
31
4B
72
90
B7
31
0B
73
08
B7
29
6B
72
29
B7
21
3B
72
62
B7
24
9B
78
25
B7
35
2B
73
48
B7
30
2B
72
54
B7
35
6B
77
51
β-g
luco
sid
ase
acti
vity
(U
/g d
ry c
ell
wei
gh
t) *B. catenulatum
*
B. infantis
* B. breve
*
B. pseudocatenulatum
*
B. adolescentis
The health image of TFPs: the role of microorganisms
-glucosidase activity: results
0 4 8 12Time (min)
kaempferol 3-O-xylosylglucoside
kaempferol 3-O-glucoside
kaempferol 7-O-glucoside
Flavonoid metabolism: selected bifidobacteria strains incubated with flavonoids extracted from “Zolfino” bean
seedsKaempferol
AFTER incubation with B7003
Kaempferol-3-0-xylosylglucoside
The health image of TFPs: the role of microorganisms
Glycosidic forms of daidzein, genistein,
glycitein, kaempferol
ChromatogramSeedling extractZolfino
landrace
0 4 8 12Time (min)
Chromatogramafter 24 h incubation
with B7003
Genistein
Glycitein
Daidzein
Kaempferol
Flavonoid metabolism: selected bifidobacteria strains incubated with flavonoids extracted from “Zolfino”
bean seedlings
The health image of TFPs: the role of microorganisms
0.00
0.05
0.10
0.15
0.20
0.25
Daidzein Genistein Glycitein Kaempferol
Deg
rad
ati
on
rate
k (
1/h
)
Metabolism of flavonoid aglycones degradation rates following incubation with selected strains
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.
Daidzein
Glycitein
Genistein
Kaempferol
Marotti I. et al. J. Agric. Food Chem. 2007, 55, 3913-3919
The health image of TFPs: the role of microorganisms
0
2
4
6
8
10
= L. fermentum
= L. buchneri
= L. kefiri
= L. helveticus
= L. plantarum
= L. bulgaricus
β-g
luco
sid
ase
acti
vity
(U
/g d
ry c
ell
wei
gh
t)
= L. lactis
L1
2 L 7 L 8 L 3 L 2 L1
1 L1
3 L 9 L1
5 L1
0 L 5 L 1 L1
4
= L. acidophilus
Studies in progress: wider screening of Lactobacillus strains
Screening of β-glucosidase activity in Lactobacillus spp.: preliminary results
The health image of TFPs: the role of microorganisms
PERSPECTIVES
Potentiality of investigated Lactobacillus strains as functional starter cultures for new fermented products facilitate bioavailability
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
The health image of TFPs: the role of microorganisms
41
Roles of TFPs against “obesity”
• dietary abundance• sedentary lifestyle
Obesity is associated with significant risks of ill health
Roles of TFPs against “obesity”
42
Overnutrition and energy density
400
800
1200
1600
0
Ene
rgy
dens
ity (
kJ 1
00 g
-1)
Des
sert
sC
hick
en
Win
gs
Frie
s (c
hip
s)C
rispy
str
ips
San
dwic
hes
Veg
eta
bles
Bis
cuit
Pop
corn
Outlet ABurger King
Des
sert
sF
ries
(chi
ps)
San
dwic
hes
Frie
d +
sid
eC
ondi
men
ts,
etc
San
dwic
hes
Bre
akf
ast
Outlet BJack in the Box
Outlet CMc Donald’s
Muf
fin/D
anis
hF
ries
(chi
ps)
Chi
cke
n M
cNu
gget
sD
ress
ings
Bre
akf
ast
Sau
ces
Des
sert
/Sha
kes
Sal
ads
San
dwic
hes
Traditional Gambian
Mea
lsS
tapl
esS
tauc
esB
rea
kfas
t
Average 1167 1087 1054 439
British diet (670)
Prentice & Jebb (2003), Obesity reviews, 4, 187-
194.
Roles of TFPs against “obesity”
43
0 400 800 1200 1600 2000
0
1000
2000
3000
4000
5000
Traditional Gambian
Market (ready dishes)
Fast foods
Energy density (kJ 100 g-1)
Wei
ght
of f
ood
eate
n (g
)Overnutrition and energy density
For assuring 8500 kJ per day
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.
Prentice & Jebb (2003), Obesity reviews, 4, 187-
194.
Roles of TFPs against “obesity”
44
Obesity predisposes the individual to the development of non-transmissible diseases
0 1 2 3 4 5 6 7
Netherlands
Sweden
Germany
Denmark
UK
France
Italy
Spain
Greece
Mortality ratios (%)
Whole cohort set Mediterranean diet
The role of traditional Mediterranean diet
Trichopoulou et al., (2005), BMJ, 330, 991-997.
• high intake of vegetables, legumes, fruit, and cereals (in the past largely unrefined);• moderate to high intake of fish;• low intake of saturated lipids but high intake of unsaturated lipids (olive oil)• moderate intake of dairy products (mostly cheese and yogurt)• low intake of meat
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
45
Perspectives and challenges of TFP sector
….. this is what the modern consumer and the modern scientist always keep in mind……
46
Perspectives and challenges of TFP sector
Personalized Nutrigenetic Food Age
18th 19th 20th 21th Century
Future
Val
ue (
Tec
hnol
ogy
+Iin
form
atio
n)
Survival food
Convenience food
Essential nutrients Engineered food Mass production
Functional food
Chemical analysis Nutraceutical Target group Health claim
Nutrigenetic food
Molecular nutrition Tailor-made personal foodHome-test kits
Harmonization of Eastern & W
estern
Health knowledge
47
Perspectives and challenges of TFP sector
Comparison
Personalized Nutrigenetic Food Age
Pathologic status Health status
TFPs
Tailor made diet
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