bio-fortification of staple foods with iron and zinc · bio-fortification of staple foods with iron...

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Bio-fortification of staple foods with iron and zinc Dr. Diego Moretti, Human Nutrition Laboratory, ETH Zürich; WFS Conference 2015

14.09.2015 Diego Moretti 1

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Food that constitutes a dominant portion of a standard diet in a given population

Wheat Rice Maize Rye Barley Potato Beans Millet Sorghum

What are staple foods


Cereals account for 52% of calories WW Africa: 60-75% Latin America: 50% Mexico: 43% US diet: 26%

Poor people eat staples

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Have the nutrient levels in grains decreased ?

3 Fischer, FAO, 2009

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Fe and Zn level in historical wheat grains

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Landraces and wilthype wheat are richer in micronutrients

Time explains ≈30% of the variation in the Fe, Zn content of wheat.

Nitrogen application has stonger effect on micronutrient content.

Monasterio and Graham, 2000, Food Nutrition Bulletin

Presenter

Presentation Notes

Has the content of minerals decreased in grains following the green revolution ?

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Wheat bio-fortification


Application of foliar Zn

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Zn localization in wheat, biofortified vs control


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Foliar Zinc application


45% ↑ Zn in endosperm, 35% ↑ Zn in bran and embryo, but at high level

Cakmak, J. Agric. Food Chem., 2010

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Phytic acid decreases with milling Fe decreases with milling Molar ratio Phytic Acid: Fe decreases with milling Fractional bioavailability increases Fe concentration decreases with extraction rate. What wheat should one eat ?

Wheat, effect of milling


Extraction rate

Con

tent

(mol

)

Fe, Zn

Phytic acid

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Wheat cultivars (Triticum dicoccoides, Esperia)

Signorell, unpublished.

Presenter

Presentation Notes

If we look at the average content of Zn in whole wheat.

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Zinc absorption study design


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Zinc absorption study design

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100% extraction rate 80% extraction rate Meal Type A B C D E F

Meal description control fortified biofortified control fortified biofortified

Final Zn [mg] 6.0 9.6 10.2 4.0 6.6 6.7 Final PA [g] 1.6 1.6 1.6 1.0 1.0 1.0 PA:Zn molar

ratio 26.4 16.6 15.4 24 15.0 14.8

FAZ [%] 9.02c ±3.42

7.73c ±2.36

8.89c ±2.79

15.94a

±6.01 11.94b

±3.25 13.32b ±4.60

Test meal composition


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Controla Fortifiedb Bio-fortifiedc

Absorbed Zn from 200 g wheat (100%) extraction

Signorell et al, 2015, unpublished

40% increase in the amount of absorbed Zn compared to conventional wheat. 1/3 of daily requirement of absorbed Zn

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120 g wheat ; 6 days a week; 5 months N=270

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Millet


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Pearl millet PA Iron PA:iron

mg/60 g flour

Regular-iron pearl millet (DG-9444) 392 ± 10 1.5 ± 0.2 22.1:1

Iron-biofortified pearl millet (ICPT8203) 511 ± 21 5.5 ± 0.6 8.2:1

Test meals – 1

Local millet paste ("pate de mil", DM 60 g) accompanied by: 110 g leafy vegetable sauce in the morning 80 g okra sauce at noon 3.7 mg Fe as 56FeSO4/post-harvest iron-fortified millet meal 0.4 mg isotopic tag dissolved in 80 g mineral water, served after half of the meal

was consumed 220 g mineral water administered at the end of the meal

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PA, phytic acid; PA:iron, phytic acid:iron molar ratio

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10.4 a

7.5 b 7.5 b

0.0

5.0

10.0

15.0

20.0

25.0

30.0

Post-harvestfortified millet

Regular millet Biofortified millet

Frac

t. Fe

abs

orpt

ion

[%]

Results – Iron absorption

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Geometric means with -1 SE and +1 SE as whiskers (Repeated measure Anova + Bonferroni corrected pairwise comparison)

1,500 a

527 b

1,125 c

0

400

800

1200

1600

2000

2400

2800

3200

3600

Post-harvestfortified millet

Regular millet Biofortified millet

Tota

l Fe

abso

rptio

n [µ

g]

P < 0.05 P < 0.01

P < 0.05 P < 0.0001

C.Cercamondi et al, J.Nutrition, 2013

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Iron bioavailability from biofortified beans

18 14.09.2015 Diego Moretti

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Fe bioavailability from Biofortified beans Iron (mg/100g)

PP (mg GA eq./100g)

PA (mg/100g) PA:Fe

Biofortified Bean 8.8 ± 0.4 570 ± 10 1320 ± 10 13:1

Control Bean 5.4 ± 0.1 450 ± 16 980 ± 30 15:1


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Low phytic acid crops

Mutations that substantially reduce PA in seeds Enhance bioavailability


(Mendoza et al. AJCN 1998)

lpa1

20

0

100

200

300

400

500

600

700

800

900

Wild type maize Low phytic acid maize

Phyt

ic a

cid

(mg/

100

g)

0

1

2

3

4

5

6

7

8

9

Iron

abso

rptio

n (%

)

Phytic acid reduction by 56% increased iron absorption 1.5 times

molar ratio PA:Fe from 16 to 8.

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LR33 mutant myo-inositol -3-phosphate synthase gene Decreased PA, matched with higher free P(i)

lpa1 ABC transporter gene transporting Phytate in seed storage

lpa2 myo inositol kinase decreased PA, higher hypophosporylated inositols

lpa3 myo inositol kinase accumulation of myo inositol

germination and emergence stress tolerance seed filling

LPA bean from mutagenesis experiments


Campion, 2009, Theor appl gen.

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LPA-280-10 myo-inositol -3-phosphate synthase gene

Beans appear to have 2 copies of the gene Expressed in the seed and one in the plasmid


Fileppi, 2010, Mol Breeding.

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Iron bioavailability from LPA-280-10

Single meal study (one 50g) Enhanced fractional absorption compared to wt beans

Multiple meal study Transient complains in a large part of the participating subjects Minimal increase in total iron absorbed.


Fe (mg/100g) PA (mg/100g) Molar ratio PA:Fe Fe absorption (%); mg

LPA 9.4 70 0.6:1 6.1; 0.372 Wt 9.2 1030 11:1 3.8; 0.235

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Multiple meal study

Bean or test meal Iron PA PA: Fe Fe absorption (%

Lpa bean, mg/100g 7.0 110 1.3:1 8.6; 0.340a

Biofortified bean, mg/100g 9.9 1208 10.4:1 7.3; 0.404a

Control bean, mg/100g 5.2 1005 16.5:1 8.0; 0.235b


Transient epigastric discomfort in participants only LPA bean Hard to cook behavior Pectin acids in the middle lamella interacting with free Ca and Mg ions in absence of PA

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Future perspectives: nutritonal research in bio-fortification

Need for sensitive nutritional assessment methods. Dietary interventions Longer trials, more power, sensitive measurements. Bio fortified foods are not therapeutic foods

Food processing paramount for biofortification Milling Phytate removal Appropriate posthavest treatement


Presenter

Presentation Notes

These are all examples how nutrition should not be seen in isolation.

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1770, 20 years

before the French Revolution (1789)

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After another poor harvest, 1770: France

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Antoine Parmentier

Potatoes were considered useful animal fodder

Growing underground Poisonous (green parts) Greatly increased caloric output per

ha. Frederic II of Prussia had ordered

the cultivation during a famine

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Hachis Parmentier or Shepherd’s Pie

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Antoine Parmentier Antoine Parmentier Pharmacist

Inspector general of health services under

Napoleon Bonaparte

Introduced the first mandatory smallpox vaccination (Pocken, Vaiolo)

Studied food preservation & refrigeration

Pioneered sugar extraction from sugar beet

1763: French parliament had banned the cultivation of the potato

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After another poor harvest: 1770: France A prize was offered for : ‘ Identifying foodstuffs capable of reducing the calamities of famine’

Parmentier won it with an elogy of the potato.

But convincing people to consume Potatoes was different than convincing his fellow scientists. Parmentier started hosting dinners with ‘ Celebrities’ cooking potato

dishes He served potatoes to Louis XVI and Marie-Antoniette Pubished recipy books with Potatoes

Was one of the first recipients of the Legion d’Honneur, by Napoleon.

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Biofortification Developed, developing countries (4) Different approaches (1) Breeding: Millet and Beans (2), phytic acid, isotopes (1). Agronomic. Foliar application of Zn (3) Urea and Fe (2), in wheat. Improved mutants : LPA bean (2) Processing

Outlook EU fortification guidelines Processing Plant enhancers of bioavailabilty ?Nicotimnamide?


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Dietary diversification

Supplementation

Food fortification and Biofortification of staple foods

Control of micronutrient deficiencies


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Zinc Bioavailability

19.11.2013 35 Hambidge et al., 2010,

Main determinants 1. Physiological factors 2. Dietary factors : phytate; zinc content in the diet

Presenter

Presentation Notes

nositol hexaphosphates and pentaphosphates are the phytate forms that exert these negative effects, whereas the lower phosphates have no or little effect on zinc absorption on can have a negative effect on zinc absorption, if given together in a supplement, whereas no effect is observed when the same amounts are present in a meal as fortificants. Cadmium, which is increasing in the environment, also inhibits zinc absorption. The amount of protein in a meal has a positive effect on zinc absorption, but individual proteins may act differently; e.g., casein has a modest inhibitory effect of zinc absorption compared with other protein sources. Amino acids, such as histidine and methionine, and other low-molecular-weight ions, such as EDTA and organic acids (e.g., citrate), are known to have a positive effect on zinc absorption and have been used for zinc supplements.

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Different forms of food iron ◦ Heme Fe Absorption of 20-30% Absorbed intact, little effect of

the diet, and less affected by Fe status.

◦ Non heme Fe: Absorption

varying between 1%-100%. Affected by dietary inhibitors Strongly affected by Fe status

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Inhibitors and enhancers of Fe abs.

◦ INHIBITORS Bind iron in the duodenum into complexes from which iron is

unavailable for absorption. ◦ Phytate, polyphenols, calcium, casein, soy protein. ◦ Whole grains, red wine, coffee, tea, cow’s milk,

legumes (soy). ◦ ENHANCERS Reduce iron, facilitate solubilisation (preventing precipitation), bind

iron in soluble complexes ◦ Ascorbic Acid (vitamin C), Muscle protein, Organic acids. ◦ Citrus fruits, certain vegetables (Broccoli, Cabbage, Chard),

meat.

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Biofortification

Agronomic approach (2) Traditional breeding (3)

Transgenic approach


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Global estimates for zinc nutrition


Wessells KR, Brown KH (2012) PLoS ONE 7(11): e50568.

Estimated prevalence of zinc deficiency globally: 17.3% Estimated substantial mortality in children each year Black RE, et al. The Lancet. Vol382(9890):427-451

Estimated % inadequate zinc intake

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Global estimates for IDA


Kassebaum NJ et al. Blood, 2014;123(5):615-624

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Bioavailability Biological efficacy

Nutritional viability


Amount of nutrient absorbed and utilized by the body Fraction of the dose (%) Absolute amount (mg)

Regular consumption improves markers of nutritional status

Presenter

Presentation Notes

By their nature, nutritional interventions have to do with small changes in nutrient level, changes take place over long term. We could use intake data, but most of the time this data is only partly considered a good predictor of nutritional efficacy because of bioavailability. Introduce phytic acid.

bio-fortification of staple foods with iron and zinc · bio-fortification of staple foods with iron...

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