Characterizing the gut (Gallus gallus) microbiota following the consumption of an iron biofortified Rwandan cream seeded carioca (Phaseolus Vulgaris L.) bean-based diet.
ABSTRACT: Biofortification is a plant breeding method that introduces increased concentrations of minerals in staple food crops (e.g., legumes, cereal grains), and has shown success in alleviating insufficient Fe intake in various human populations. Unlike other strategies utilized to alleviate Fe deficiency, studies of the gut microbiota in the context of Fe biofortification have not yet been reported, although the consumption of Fe biofortified staple food crops has increased significantly over time. Hence, in this study, we performed a 6-week feeding trial in Gallus gallus (n = 14), aimed to investigate the alterations in the gut microbiome following administration of an Fe biofortified bean-based diet (biofortified, BFe) versus a bean based diet with poorly-bioavailable Fe (standard, SFe). Cream seeded carioca bean based diets were designed in an identical fashion to those used in a recent human clinical trial of Fe biofortified beans in Rwanda. We hypothesized that the different dietary Fe contents in the beans based diets will alter the composition and function of the intestinal microbiome. The primary outcomes were changes in the gut microbiome composition and function analyzed by 16S rRNA gene sequencing. We observed no significant changes in phylogenetic diversity between groups. There were significant differences in the composition of the microbiota between groups, with the BFe group harboring fewer taxa participating in bacterial Fe uptake, increased abundance of bacteria involved in phenolic catabolism, and increased abundance of beneficial butyrate-producing bacteria. Additionally, depletion of key bacterial pathways responsible for bacterial viability and Fe uptake suggest that improvements in Fe bioavailability, in addition to increases in Fe-polyphenol and Fe-phytate complexes due to biofortification, led to decreased concentrations of cecal Fe available for bacterial utilization. Our findings demonstrate that Fe biofortification may improve Fe status without negatively altering the structure and function of the gut microbiota, as is observed with other nutritional methods of Fe supplementation. These results may be used to further improve the efficacy and safety of future biofortification efforts in eradicating global Fe deficiency.
Project description:Iron (Fe) deficiency is a highly prevalent micronutrient insufficiency predominantly caused by a lack of bioavailable Fe from the diet. The consumption of beans as a major food crop in some populations suffering from Fe deficiency is relatively high. Therefore, our objective was to determine whether a biofortified variety of cream seeded carioca bean (Phaseolus vulgaris L.) could provide more bioavailable-Fe than a standard variety using in-vivo (broiler chicken, Gallus gallus) and in-vitro (Caco-2 cell) models. Studies were conducted under conditions designed to mimic the actual human feeding protocol. Two carioca-beans, a standard (G4825; 58 ?g Fe/g) and a biofortified (SMC; 106 ?g Fe/g), were utilized. Diets were formulated to meet the nutrient requirements of Gallus gallus except for Fe (33.7 and 48.7 ?g Fe/g, standard and biofortified diets, respectively). In-vitro observations indicated that more bioavailable-Fe was present in the biofortified beans and diet (P<0.05). In-vivo, improvements in Fe-status were observed in the biofortified bean treatment, as indicated by the increased total-body-Hemoglobin-Fe, and hepatic Fe-concentration (P<0.05). Also, DMT-1 mRNA-expression was increased in the standard bean treatment (P<0.05), indicating an upregulation of absorption to compensate for less bioavailable-Fe. These results demonstrate that the biofortified beans provided more bioavailable Fe; however, the in vitro results revealed that ferritin formation values were relatively low. Such observations are indicative of the presence of high levels of polyphenols and phytate that inhibit Fe absorption. Indeed, we identified higher levels of phytate and quercetin 3-glucoside in the Fe biofortified bean variety. Our results indicate that the biofortified bean line was able to moderately improve Fe-status, and that concurrent increase in the concentration of phytate and polyphenols in beans may limit the benefit of increased Fe-concentration. Therefore, specific targeting of such compounds during the breeding process may yield improved dietary Fe-bioavailability. Our findings are in agreement with the human efficacy trial that demonstrated that the biofortified carioca beans improved the Fe-status of Rwandan women. We suggest the utilization of these in vitro and in vivo screening tools to guide studies aimed to develop and evaluate biofortified staple food crops. This approach has the potential to more effectively utilize research funds and provides a means to monitor the nutritional quality of the Fe-biofortified crops once released to farmers.
Project description:The common dry bean (Phaseolus vulgaris L.) is a globally produced pulse crop and an important source of micronutrients for millions of people across Latin America and Africa. Many of the preferred black and red seed types in these regions have seed coat polyphenols that inhibit the absorption of iron. Yellow beans are distinct from other market classes because they accumulate the antioxidant kaempferol 3-glucoside in their seed coats. Due to their fast cooking tendencies, yellow beans are often marketed at premium prices in the same geographical regions where dietary iron deficiency is a major health concern. Hence, this study compared the iron bioavailability of three faster cooking yellow beans with contrasting seed coat colors from Africa (Manteca, Amarillo, and Njano) to slower cooking white and red kidney commercial varieties. Iron status and iron bioavailability was assessed by the capacity of a bean based diet to generate and maintain total body hemoglobin iron (Hb-Fe) during a 6 week in vivo (Gallus gallus) feeding trial. Over the course of the experiment, animals fed yellow bean diets had significantly (p ≤ 0.05) higher Hb-Fe than animals fed the white or red kidney bean diet. This study shows that the Manteca yellow bean possess a rare combination of biochemical traits that result in faster cooking times and improved iron bioavailability. The Manteca yellow bean is worthy of germplasm enhancement to address iron deficiency in regions where beans are consumed as a dietary staple.
Project description:Biofortification is an effective method to improve the nutritional content of crops and nutritional intake. Breeding for higher micronutrient mineral content in beans is correlated with an increase in phytic acid, a main inhibitor of mineral absorption in humans. Low phytic acid (lpa) beans have a 90% lower phytic acid content compared to conventional beans. This is the first study to investigate mineral and total phytic acid retention after preparing common household recipes from conventional, biofortified and lpa beans. Mineral retention was determined for two conventional, three biofortified and two lpa bean genotypes. Treatments included soaking, boiling (boiled beans) and refrying (bean paste). The average true retention of iron after boiling was 77.2-91.3%; for zinc 41.2-84.0%; and for phytic acid 49.9-85.9%. Soaking led to a significant decrease in zinc and total phytic acid after boiling and refrying, whereas for iron no significant differences were found. lpa beans did not exhibit a consistent pattern of difference in iron and phytic acid retention compared to the other groups of beans. However, lpa beans had a significantly lower retention of zinc compared to conventional and biofortified varieties (p < 0.05). More research is needed to understand the underlying factors responsible for the differences in retention between the groups of beans, especially the low retention of zinc. Combining the lpa and biofortification traits could further improve the nutritional benefits of biofortified beans, by decreasing the phytic acid:iron and zinc ratio in beans.
Project description:BACKGROUND:Iron-biofortified staple foods can improve iron status and resolve iron deficiency. However, whether improved iron status from iron biofortification can improve physical performance remains unclear. OBJECTIVE:This study aimed to examine whether changes in iron status from an iron-biofortified bean intervention affect work efficiency. METHODS:A total of 125 iron-depleted (ferritin <20 ?g/L) female Rwandan university students (18-26 y) were selected from a larger sample randomly assigned to consume iron-biofortified beans (Fe-Bean; 86.1 mg Fe/kg) or conventional beans (control: 50.6 mg Fe/kg) twice daily for 18 wk (average of 314 g beans consumed/d). Blood biomarkers of iron status (primary outcome) and physical work efficiency (secondary outcome) were measured before and after the intervention. Work performed was assessed during 5-min steady-state periods at 0-, 25-, and 40-W workloads using a mechanically braked cycle ergometer. Work efficiency was calculated at 25 W and 40 W as the work accomplished divided by the energy expended at that workload above that expended at 0 W. General linear models were used to evaluate the relation between changes in iron status biomarkers and work efficiency. RESULTS:The Fe-Bean intervention had significant positive effects on hemoglobin, serum ferritin, and body iron stores but did not affect work efficiency. However, 18-wk change in hemoglobin was positively related to work efficiency at 40 W in the full sample (n = 119; estimate: 0.24 g/L; 95% CI: 0.01, 0.48 g/L; P = 0.044) and among women who were anemic (hemoglobin <120 g/L) at baseline (n = 43; estimate: 0.64 g/L; 95% CI: 0.05, 1.23 g/L; P = 0.036). Among women who were nonanemic at baseline, change in serum ferritin was positively related to change in work efficiency at 40 W (n = 60; estimate: 0.50 ?g/L; 95% CI: 0.06, 0.95 ?g/L; P = 0.027). CONCLUSIONS:Increasing iron status during an iron-biofortified bean feeding trial improves work efficiency in iron-depleted, sedentary women. This trial was registered at clinicaltrials.gov as NCT01594359.
Project description:Staple food crops tend to be low in micronutrients; therefore, individuals whose diets rely heavily on them can suffer from micronutrient deficiency. Biofortification addresses this issue through the breeding of staple crops that are micronutrient-dense and high yielding. One such crop is iron-biofortified beans. Ten iron-biofortified bean varieties were released between 2010 and 2012 in Rwanda, a country with high rates of bean production and consumption, to address iron deficiency. This study evaluates the effect of the most widely adopted of these varieties, RWR2245, on household yield, land cultivated under beans, bean consumption, purchases, and sales. Because the adoption decision could be endogenous, we use a control function approach to quantify the impacts of adoption. RWR2245 provides a yield gain of 20%-49% over traditional bush bean varieties. In our preferred model specification, we find that over a 12-month period, growing RWR2245 for at least one out of two annual growing seasons increases the length of time beans are consumed from own production by 0.64 months (19-20 days), reduces the length of time beans are purchased for consumption by 0.73 months (22-23 days), and increases the probability of selling beans by 12%. Adoption can thus improve household nutrition via two channels: primarily by increasing iron intake via substituting biofortified harvested beans for less nutrient-dense beans from the market, and additionally by increasing household income that can be spent on nutritious foods through the reduction in bean purchases and increased likelihood of selling beans. Moreover, the sale of iron-biofortified beans implies the availability of iron-dense food in markets, also benefiting households that purchase beans. These findings are promising for the continued adoption of iron-biofortified beans in Rwanda and elsewhere and provide evidence that biofortified crops are an effective investment for nutrition, food security, and poverty reduction.
Project description:In many poorer parts of the world, biofortification is a strategy that increases the concentration of target nutrients in staple food crops, mainly by genetic manipulation, to alleviate prevalent nutrient deficiencies. We reviewed the (i) prevalence of vitamin A, iron (Fe) and zinc (Zn) deficiencies; (ii) availability of vitamin A, iron and Zn biofortified crops, and their acceptability in South Africa. The incidence of vitamin A and iron deficiency among children below five years old is 43.6% and 11%, respectively, while the risk of Zn deficiency is 45.3% among children aged 1 to 9 years. Despite several strategies being implemented to address the problem, including supplementation and commercial fortification, the prevalence of micronutrient deficiencies is still high. Biofortification has resulted in the large-scale availability of ?carotene-rich orange-fleshed sweet potatoes (OFSP), while provitamin A biofortified maize and Zn and/or iron biofortified common beans are at development stages. Agronomic biofortification is being investigated to enhance yields and concentrations of target nutrients in crops grown in agriculturally marginal environments. The consumer acceptability of OFSP and provitamin A biofortified maize were higher among children compared to adults. Accelerating the development of other biofortified staple crops to increase their availability, especially to the target population groups, is essential. Nutrition education should be integrated with community health programmes to improve the consumption of the biofortified crops, coupled with further research to develop suitable recipes/formulations for biofortified foods.
Project description:Iron deficiency is a major public health problem worldwide, with the highest burden among children. The objective of this randomized efficacy feeding trial was to determine the effects of consuming iron-biofortified beans (Fe-Beans) on the iron status in children, compared to control beans (Control-Beans). A cluster-randomized trial of biofortified beans (Phaseolus vulgaris L), bred to enhance iron content, was conducted over 6 months. The participants were school-aged children (n = 574; 5?12 years), attending 20 rural public boarding schools in the Mexican state of Oaxaca. Double-blind randomization was conducted at the school level; 20 schools were randomized to receive either Fe-Beans (n = 10 schools, n = 304 students) or Control-Beans (n = 10 schools, n = 366 students). School administrators, children, and research and laboratory staff were blinded to the intervention group. Iron status (hemoglobin (Hb), serum ferritin (SF), soluble transferrin receptor (sTfR), total body iron (TBI), inflammatory biomarkers C-reactive protein (CRP) and -1-acid glycoprotein (AGP)), and anthropometric indices for individuals were evaluated at the enrollment and at the end of the trial. The hemoglobin concentrations were adjusted for altitude, and anemia was defined in accordance with age-specific World Health Organization (WHO) criteria (i.e., Hb <115 g/L for <12 years and Hb <120 g/L for 12 years). Serum ferritin concentrations were adjusted for inflammation using BRINDA methods, and iron deficiency was defined as serum ferritin at less than 15.0 µg/L. Total body iron was calculated using Cook's equation. Mixed models were used to examine the effects of Fe-Beans on hematological outcomes, compared to Control-Beans, adjusting for the baseline indicator, with school as a random effect. An analysis was conducted in 10 schools (n = 269 students) in the Fe-Beans group and in 10 schools (n = 305 students) in the Control-Beans group that completed the follow-up. At baseline, 17.8% of the children were anemic and 11.3% were iron deficient (15.9%, BRINDA-adjusted). A total of 6.3% of children had elevated CRP (>5.0 mg/L), and 11.6% had elevated AGP (>1.0 g/L) concentrations at baseline. During the 104 days when feeding was monitored, the total mean individual iron intake from the study beans (Fe-bean group) was 504 mg (IQR: 352, 616) over 68 mean feeding days, and 295 mg (IQR: 197, 341) over 67 mean feeding days in the control group (p < 0.01). During the cluster-randomized efficacy trial, indicators of iron status, including hemoglobin, serum ferritin, soluble transferrin receptor, and total body iron concentrations improved from the baseline to endline (6 months) in both the intervention and control groups. However, Fe-Beans did not significantly improve the iron status indicators, compared to Control-Beans. Similarly, there were no significant effects of Fe-Beans on dichotomous outcomes, including anemia and iron deficiency, compared to Control-Beans. In this 6-month cluster-randomized efficacy trial of iron-biofortified beans in school children in Mexico, indicators of iron status improved in both the intervention and control groups. However, there were no significant effects of Fe-Beans on iron biomarkers, compared to Control-Beans. This trial was registered at clinicaltrials.gov as NCT03835377.
Project description:Food plants biofortification for micronutrients is a tool for the nutritional value improvement of food. Soilless cultivation systems, with the optimal control of plant nutrition, represent a potential effective technique to increase the beneficial element content in plant tissues. Silicon (Si), which proper intake is recently recommended for its beneficial effects on bone health, presents good absorption in intestinal tract from green bean, a high-value vegetable crop. In this study we aimed to obtain Si biofortified green bean pods by using a Si-enriched nutrient solution in soilless system conditions, and to assess the influence of boiling and steaming cooking methods on Si content, color parameters and Si bioaccessibility (by using an in vitro digestion process) of pods. The Si concentration of pods was almost tripled as a result of the biofortification process, while the overall crop performance was not negatively influenced. The Si content of biofortified pods was higher than unbiofortified also after cooking, despite the cooking method used. Silicon bioaccessibility in cooked pods was more than tripled as a result of biofortification, while the process did not affect the visual quality of the product. Our results demonstrated that soilless cultivation can be successfully used for green bean Si biofortification.
Project description:Micronutrient deficiencies, also known as hidden hunger, affect two billion people worldwide, curtailing their ability to lead healthy, productive lives. Biofortified staple crops, bred to be rich in micronutrient content, are a cost-effective and scalable solution to alleviating micronutrient deficiency, particularly among rural households who consume what they produce. Delivery of biofortified planting material in Rwanda began in 2012, and it is important to learn from the efforts undertaken to date to inform the design of higher impact - lower cost delivery strategies for scaling up these crops. In this paper, we use a nationally representative household survey of bean producers and delivery data from seven consecutive seasons and apply duration analysis to estimate the impact of different delivery approaches on household time to adoption, disadoption and readoption of iron-biofortified beans in Rwanda. Proximity to formal delivery via sales of small packets of planting material quickens adoption and readoption, while delivery of larger quantities of planting material to small-scale producers within a village slows disadoption of iron-biofortified beans. Informal dissemination within social networks and access to extension are also major drivers of rapid adoption. In addition, households whose main decision maker for bean production is a woman, has some formal education, and more years of experience growing beans disadopt iron-biofortified beans more slowly than other households. These findings provide evidence that current efforts to promote iron-biofortified crops have been successful and are expected to inform future development of sustainable and cost-effective delivery models for biofortified crops in Rwanda and elsewhere.
Project description:Mineral deficiency is worldwide one of the major problems associated with human health, and biofortification through breeding is considered an important strategy to improve the nutritional content of staple food in countries that face this problem. The assessment of genetic variability for seed nutrient contents is a first step in the development of a biofortified crop. From the germplasm bank IDR-IAPAR-EMATER, a set of 1,512 common bean accessions, consisting of local and commercial varieties and improved lines, was analyzed. High variability among the accessions was observed for all evaluated nutrient contents (P, K, Ca, Mg, Cu, Zn, Mn, Fe and S and protein). In the mean, the contents of the carioca and black market groups (Mesoamerican gene pool), were around 7% higher for the minerals Ca, Cu, Mn and Fe and between 2-4% higher for P, K, Mg and Zn than in the other groups with Mesoamerican and Andean common bean. Few differences were observed among the Mesoamerican accessions that belong to the carioca and black commercial groups. Wide variability was observed among the evaluated genotypes, and the concentrations of the best accessions exceeded the overall mean by 14-28%. Due to the high variability in the evaluated accessions, these results may contribute to the selection of promising parents for the establishment of mating blocks. The nutritional contents of many of the improved lines evaluated in this study were higher than those of the commercial cultivars, indicating the possibility of developing new biofortified cultivars.