Fabiana F. De Moura

Bioavailability of iron, zinc, and provitamin A carotenoids in biofortified staple crops

International research efforts, including those funded by HarvestPlus, a Challenge Program of the Consultative Group on International Agricultural Research (CGIAR), are focusing on conventional plant breeding to biofortify staple crops such as maize, rice, cassava, beans, wheat, sweet potatoes, and pearl millet to increase the concentrations of micronutrients that are commonly deficient in specific population groups of developing countries. The bioavailability of micronutrients in unfortified staple crops in developing regions is typically low, which raises questions about the efficacy of these crops to improve population micronutrient status. This review of recent studies of biofortified crops aims to assess the micronutrient bioavailability of biofortified staple crops in order to derive lessons that may help direct plant breeding and to infer the potential efficacy of food-based nutrition interventions. Although reducing the amounts of antinutrients and the conduction of food processing generally increases the bioavailability of micronutrients, antinutrients still possess important benefits, and food processing results in micronutrient loss. In general, biofortified foods with relatively higher micronutrient density have higher total absorption rates than nonbiofortified varieties. Thus, evidence supports the focus on efforts to breed plants with increased micronutrient concentrations in order to decrease the influence of inhibitors and to offset losses from processing.

Retention of Provitamin A Carotenoids in Staple Crops Targeted for Biofortification in Africa: Cassava, Maize and Sweet Potato

HarvestPlus, part of the Consultative Group on Internation Agriculture research (CGIAR) Program on Agriculture for Nutrition and Health (A4NH) uses conventional plant breeding techniques to develop staple food crops that are rich in micronutrients, a food-based approach to reduce micronutrient malnutrition known as biofortification. The nutritional breeding targets are established based on the food intake of target populations, nutrient losses during storage and processing and bioavailability. This review collates the evidence on the retention of provitamin A carotenoid (pVAC) after processing, cooking, and storing of the staple crops targeted for pVAC biofortification: cassava, maize, and sweet potato. Sun drying was more detrimental to the pVAC levels (27–56% retention) in cassava than shade (59%) or oven (55–91%) drying, while the pVAC retention levels (66–96%) in sweet potato were not significantly different among the various drying methods. Overall, boiling and steaming had higher pVAC retention (80–98%) compared to baking (30–70%) and frying (18–54%). Gari, the most frequently consumed form of cassava in West Africa had the lowest pVAC retention (10–30%). The pVAC retention of maize grain and cassava and sweet potato flour reached levels as low as 20% after 1–4 months of storage and was highly dependent on genotype. Therefore, we recommend that an evaluation of the pVAC degradation rate among different genotypes be performed before a high pVAC crop is promoted.

Retention during Processing and Bioaccessibility of β-Carotene in High β-Carotene Transgenic Cassava Root

Cassava is a root crop that serves as a primary caloric source for many African communities despite its low content of β-carotene (βC). Carotenoid content of roots from wild type (WT) and three transgenic lines with high βC were compared after cooking and preparation of nonfermented and fermented flours according to traditional African methods. The various methods of processing all decreased βC content per gram dry weight regardless of genotype. The greatest loss of βC occurred during preparation of gari (dry fermentation followed by roasting) from WT and transgenic lines. The quantities of βC in cooked transgenic cassava root that partitioned into mixed micelles during in vitro digestion and transported into Caco-2 cells were significantly greater than those for identically processed WT root. These results suggest that transgenic high βC cassava will provide individuals with greater quantities of bioaccessible βC.

Bioaccessibility of carotenoids from transgenic provitamin A biofortified sorghum.

Biofortified sorghum (Sorghum bicolor (L.) Moench) lines are being developed to target vitamin A deficiency in Sub-Saharan Africa, but the delivery of provitamin A carotenoids from such diverse germplasms has not been evaluated. The purpose of this study was to screen vectors and independent transgenic events for the bioaccessibility of provitamin A carotenoids using an in vitro digestion model. The germplasm background and transgenic sorghum contained 1.0-1.5 and 3.3-14.0 μg/g β-carotene equivalents on a dry weight basis (DW), respectively. Test porridges made from milled transgenic sorghum contained up to 250 μg of β-carotene equivalents per 100 g of porridge on a fresh weight basis (FW). Micellarization efficiency of all-trans-β-carotene was lower (p < 0.05) from transgenic sorghum (1-5%) than from null/nontransgenic sorghum (6-11%) but not different between vector constructs. Carotenoid bioaccessibility was significantly improved (p < 0.05) by increasing the amount of coformulated lipid in test porridges from 5% w/w to 10% w/w. Transgenic sorghum event Homo188-A contained the greatest bioaccessible β-carotene content, with a 4-8-fold increase from null/nontransgenic sorghum. While the bioavailability and bioconversion of provitamin A carotenoids from these grains must be confirmed in vivo, these data support the notion that biofortification of sorghum can enhance total and bioaccessible provitamin A carotenoid levels.

Effect of phytate reduction of sorghum, through genetic modification, on iron and zinc availability as assessed by an in vitro dialysability bioaccessibility assay, Caco-2 cell uptake assay, and suckling rat pup absorption model

Improved iron and zinc availability from sorghum, a commonly consumed staple, will benefit many malnourished communities in rural Africa burdened with high prevalence of iron and zinc deficiency. This research compared the effect of genetic phytate reduction in sorghum on iron and zinc bioaccessibility and uptake measured by in vitro dialysability and Caco-2 cell uptake assays to that of iron and zinc absorption measured by a suckling rat pup model. The phytate reduction (80-86%) in these sorghums significantly increased zinc availability. The Caco-2 cell method, but not the dialysability assay, proved useful in estimating zinc absorption. The measured increase in iron availability differed between the methods, possibly due to the effect of varying mineral (Ca, Fe, Zn, P) contents of the sorghums. This effect was most prominent in the iron uptake results. More research is needed to determine the effect of naturally occurring variations in mineral contents of sorghum on the iron uptake by Caco-2 cells.

Carotenoid Retention of Biofortified Provitamin A Maize (Zea mays L.) after Zambian Traditional Methods of Milling, Cooking and Storage

Provitamin A biofortified maize hybrids were developed to target vitamin A deficient populations in Africa. The purpose of this study was to evaluate the degradation of carotenoids after milling, cooking, and storage among biofortified varieties released in Zambia. The biofortified maize hybrids contained 7.5 to 10.3 μg/g dry weight (DW) of provitamin A as measured by β-carotene equivalents (BCE). There was virtually no degradation due to milling. The BCE retention was also high (>100%) for most genotypes when the maize was cooked into thick (nshima) and thin porridge, but showed a lower BCE retention (53-98%) when cooked into samp (dehulled kernels). Most of the degradation occurred in the first 15 days of storage of the maize as kernels and ears (BCE retention 52-56%) which then stabilized, remaining between 30% and 33% of BCE after six months of storage. In conclusion, most of the provitamin A degradation in biofortified maize hybrids occurred during storage compared with cooking and the magnitude of this effect varied among genotypes.

Seasonal differences in food and nutrient intakes among young children and their mothers in rural Burkina Faso

It is important to understand and account for seasonal variation in food and nutrient intakes when planning interventions to combat micronutrient deficiencies in resource-poor settings. The objective of the present study was to quantify food and nutrient intakes and assess the adequacy of micronutrient intakes among young children and their mothers during the lean and post-harvest (PH) seasons in rural Burkina Faso. We quantified food intakes by 24-h recall in a representative sample of 480 children aged 36–59 months and their mothers in two provinces in Western Burkina Faso. We calculated the probability of adequacy (PA) of usual intakes of ten micronutrients and an overall mean PA (MPA). Seasonal changes in nutrient intakes and PA were assessed by mixed linear regression and non-parametric tests, respectively. Energy intakes did not differ significantly between seasons for women or children, although the womens intakes were slightly higher in the PH season. Most of the micronutrient intakes were significantly higher in the PH season, with the exception of vitamin A which was lower and vitamin B12 and Zn which were similar across seasons. MPA increased significantly across seasons, from 0·26 to 0·37 for women and from 0·43 to 0·52 for children. PA of Ca, vitamin C, folate and vitamin B12 were very low. Staple grains and vegetables were major sources of micronutrients but intakes were not sufficient to meet nutrient needs for the majority of the subjects. Food-based strategies are needed to increase micronutrient intakes of women and children in Burkina Faso.

Impact of genotype and cooking style on the content, retention, and bioacessibility of β-carotene in biofortified cassava (Manihot esculenta Crantz) conventionally bred in Brazil.

Biofortification is a strategy for decreasing micronutrient deficiencies in vulnerable populations by increasing nutrient density in staple food crops. Roots from five varieties of cassava biofortified with β-carotene (βC), three parental accessions, and one variety of commonly consumed white cassava from Brazil were investigated. Roots from biofortified varieties contained up to 23-fold higher βC than white cassava, and the additional complement of βC was primarily the all-trans isomer. At least 68% of βC per gram fresh weight was retained after boiling or boiling and briefly frying. Micellarization of βC during simulated digestion of fried root exceeded that of boiled root. Apical uptake of all-trans-βC from mixed micelles by Caco-2 cells was affected by an interaction between variety and cooking style. These results suggest that Brazilian cassava biofortified with βC has the potential to reduce vitamin A deficiency without requiring major changes in local and ethnic styles of home cooking.

Cassava Intake and Vitamin A Status among Women and Preschool Children in Akwa-Ibom, Nigeria

Background As part of the HarvestPlus provitamin A-biofortified cassava program in Nigeria we conducted a survey to determine the cassava intake and prevalence of vitamin A deficiency among children 6-59 months and women of childbearing age in the state of Akwa Ibom. Methods A cluster-randomized cross-sectional survey was conducted in 2011 in Akwa Ibom, Nigeria. The usual food and nutrient intakes were estimated using a multi-pass 24-hour recall with repeated recall on a subsample. Blood samples of children and women were collected to analyze for serum retinol, serum ferritin, and acute phase proteins as indicators of infection. Vitamin A deficiency was defined as serum retinol <0.70 μmol/L adjusted for infection. Results A total of 587 households of a mother-child dyad participated in the dietary intake assessment. Cassava was very widely consumed in Akwa Ibom, mainly as gari or foofoo. Daily cassava consumption frequency was 92% and 95% among children and women, respectively. Mean (±SD) cassava intake (expressed as raw fresh weight) was 348 ± 317 grams/day among children and 940 ± 777 grams/day among women. Intakes of most micronutrients appeared to be adequate with the exception of calcium. Median vitamin A intake was very high both for children (1038 μg RAE/day) and women (2441 μg RAE/day). Red palm oil and dark green leafy vegetables were the main sources of vitamin A in the diet, with red palm oil alone contributing almost 60% of vitamin A intake in women and children. Prevalence of vitamin A deficiency ranged from moderate (16.9 %) among children to virtually non-existent (3.4 %) among women. Conclusion Consumption of cassava and vitamin A intake was high among women and children in Akwa Ibom with a prevalence of vitamin A deficiency ranging from moderate in children to non-existent among women. The provitamin A biofortified cassava and other vitamin A interventions should focus dissemination in states where red palm oil is not widely consumed.

Biofortified β-carotene rice improves vitamin A intake and reduces the prevalence of inadequacy among women and young children in a simulated analysis in Bangladesh, Indonesia, and the Philippines

Background: Vitamin A deficiency continues to be a major public health problem affecting developing countries where people eat mostly rice as a staple food. In Asia, rice provides up to 80% of the total daily energy intake. Objective: We used existing data sets from Bangladesh, Indonesia, and the Philippines, where dietary intakes have been quantified at the individual level to 1) determine the rice and vitamin A intake in nonpregnant, nonlactating women of reproductive age and in nonbreastfed children 1–3 y old and 2) simulate the amount of change that could be achieved in the prevalence of inadequate intake of vitamin A if rice biofortified with β-carotene were consumed instead of the rice consumed at present. Design: We considered a range of 4–20 parts per million (ppm) of β-carotene content and 10–70% substitution levels for the biofortified rice. Software was used to estimate usual rice and vitamin A intake for the simulation analyses. Results: In an analysis by country, the substitution of biofortified rice for white rice in the optimistic scenario (20 ppm and 70% substitution) decreased the prevalence of vitamin A inadequacy from baseline 78% in women and 71% in children in Bangladesh. In Indonesia and the Philippines, the prevalence of inadequacy fell by 55–60% in women and dropped by nearly 30% in children from baseline. Conclusions: The results of the simulation analysis were striking in that even low substitution levels and modest increases in the β-carotene of rice produced a meaningful decrease in the prevalence of inadequate intake of vitamin A. Increasing the substitution levels had a greater impact than increasing the β-carotene content by >12 ppm.