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

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

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

Have the nutrient levels in grains decreased ?

3 Fischer, FAO, 2009

Fe and Zn level in historical wheat grains

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

Wheat bio-fortification

Application of foliar Zn

Zn localization in wheat, biofortified vs control

Foliar Zinc application

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

Cakmak, J. Agric. Food Chem., 2010

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

Fe, Zn

Phytic acid

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Wheat cultivars (Triticum dicoccoides, Esperia)

Signorell, unpublished.

Zinc absorption study design

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Zinc absorption study design

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Signorell et al, 2015, unpublished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

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

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

10.4 a

7.5 b 7.5 b

Post-harvestfortified millet

Regular millet Biofortified millet

Results – Iron absorption

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

1,500 a

1,125 c

Post-harvestfortified millet

Regular millet Biofortified millet

P < 0.05 P < 0.01

P < 0.05 P < 0.0001

C.Cercamondi et al, J.Nutrition, 2013

Iron bioavailability from biofortified beans

18 14.09.2015 Diego Moretti

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

Low phytic acid crops

Mutations that substantially reduce PA in seeds Enhance bioavailability

(Mendoza et al. AJCN 1998)

Wild type maize Low phytic acid maize

Phytic acid reduction by 56% increased iron absorption 1.5 times

molar ratio PA:Fe from 16 to 8.

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.

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.

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

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

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

1770, 20 years

before the French Revolution (1789)

After another poor harvest, 1770: France

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

Hachis Parmentier or Shepherd’s Pie

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

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.

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?

Dietary diversification

Supplementation

Food fortification and Biofortification of staple foods

Control of micronutrient deficiencies

Zinc Bioavailability

19.11.2013 35 Hambidge et al., 2010,

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

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

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),

Biofortification

Agronomic approach (2) Traditional breeding (3)

Transgenic approach

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

Global estimates for IDA

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

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

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

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