traditional food: functional properties and perspectives diana di gioia 1 department of...

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


8


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)


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

http://organic.com.au/images/logos/logo_QualityLowInputFood.gif


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



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)



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


http://images.google.it/imgres?imgurl=http://oncologiaesalute.files.wordpress.com/2008/02/bifidobacterium.jpg&imgrefurl=http://oncologiaesalute.wordpress.com/2008/02/17/i-probiotici-sono-letali-in-caso-di-pancreatite-acuta/&h=489&w=574&sz=36&hl=it&start=2&um=1&tbnid=8BFgYsLQy5eYNM:&tbnh=114&tbnw=134&prev=/images%3Fq%3Dprobiotici%26um%3D1%26hl%3Dit%26sa%3DN

15

Relevance of LAB in different fermented food products

Wine BeerCider

Sauerkraut

Fermented milk

Cheese Fermentedsausages


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


18


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

http://www.ifv.life.ku.dk/upload/ifv/food%20micro/img_food_micro/blue%20patcom%20logo%20+%20slogan%20short%20line.jpg

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


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


Selection of protective and probiotic cultures of LAB and bifidobacteria able to inhibit Campylobacter jejuni strains

http://images.google.it/imgres?imgurl=http://upload.wikimedia.org/wikipedia/commons/thumb/d/df/ARS_Campylobacter_jejuni.jpg/450px-ARS_Campylobacter_jejuni.jpg&imgrefurl=http://commons.wikimedia.org/wiki/Image:ARS_Campylobacter_jejuni.jpg&h=600&w=450&sz=80&hl=it&start=3&um=1&tbnid=o-FGhz8eq61w1M:&tbnh=135&tbnw=101&prev=/images%3Fq%3DCampylobacter%2Bjejuni%26um%3D1%26hl%3Dit%26sa%3DN


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


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


Campylobacter jejuni CIP 70.1

Campylobacter jejuni LMG 8842

Campylobacter jejuni 221/05

L. pentosus PCA 227 - + -

L. plantarum PCA 236

- + -


- + +


- + +

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


• 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%


• 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



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


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)



28


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


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


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)


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


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.


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


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


-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


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


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


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


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


41

Roles of TFPs against “obesity”

• dietary abundance• sedentary lifestyle

Obesity is associated with significant risks of ill health


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.


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.


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


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


Comparison

Personalized Nutrigenetic Food Age

Pathologic status Health status

TFPs

Tailor made diet

48

Thank you

traditional food: functional properties and perspectives diana di gioia 1 department of...

Documents

role of tfps

traditional ingredients

health image of tfps

production slide

functional properties

technology slide

role of plant ingredients

obesity slide