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
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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
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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
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Wheat bio-fortification
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Application of foliar Zn
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Zn localization in wheat, biofortified vs control
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Foliar Zinc application
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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
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Extraction rate
Con
tent
(mol
)
Fe, Zn
Phytic acid
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Wheat cultivars (Triticum dicoccoides, Esperia)
Signorell, unpublished.
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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
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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
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(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
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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
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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.
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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
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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
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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
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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
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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
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Kassebaum NJ et al. Blood, 2014;123(5):615-624
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Bioavailability Biological efficacy
Nutritional viability
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Amount of nutrient absorbed and utilized by the body Fraction of the dose (%) Absolute amount (mg)
Regular consumption improves markers of nutritional status
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