Maintenance of glucose homeostasis during late-term embryonic development is dependent upon the amount of glucose held in reserve primarily in the form of glycogen in the liver and upon the degree of glucose generated by gluconeogenesis from protein first mobilized from amnion albumen and then from muscle. Insufficient glycogen and albumen will force the embryo to mobilize more muscle protein toward gluconeogenesis, thus restricting growth of the late-term embryo and hatchling. We hypothesize that administration of available carbohydrates to the amnion will improve glycogen reserves and spare muscle protein mobilization for gluconeogenesis during late-term embryonic and posthatch neonatal development. Our hypothesis was tested by comparing BW gain, liver glycogen reserves, and muscle weight of in ovo fed and control embryos during last days of embryonic incubation until 25 d after hatching. We examined, using 600 birds from 2 different strains of commercial boilers, body and muscle weights and glycogen reserves following feeding embryos at d 17.5 of incubation with a solution containing maltose, sucrose, dextrin, and beta-hydroxy-beta-methylbutyrate (HMB). Providing carbohydrates and HMB to late-term embryos increased hatching weights by 5 to 6% over controls, improved liver glycogen by 2- to 5-fold, and elevated relative breast muscle size by 6 to 8%. These weight advantages were sustained through the end of the experiments at 25 d of age. It is reasonable to assume that the elevated glycogen levels in the in ovo treatment reduce the need to produce glucose via gluconeogenesis and, therefore, contribute to less use of muscle protein and hence a greater percentage of pectoral muscle weight in the in ovo birds.

The use of engineered nanoparticles in food and pharmaceuticals is expected to increase, but the impact of chronic oral exposure to nanoparticles on human health remains unknown. Here, we show that chronic and acute oral exposure to polystyrene nanoparticles can influence iron uptake and iron transport in an in vitro model of the intestinal epithelium and an in vivo chicken intestinal loop model. Intestinal cells that are exposed to high doses of nanoparticles showed increased iron transport due to nanoparticle disruption of the cell membrane. Chickens acutely exposed to carboxylated particles (50 nm in diameter) had a lower iron absorption than unexposed or chronically exposed birds. Chronic exposure caused remodelling of the intestinal villi, which increased the surface area available for iron absorption. The agreement between the in vitro and in vivo results suggests that our in vitro intestinal epithelium model is potentially useful for toxicology studies.

Early development of the digestive tract is crucial for achieving maximal growth and development of chickens. Because the late-term embryo naturally consumes the amniotic fluids, insertion of a nutrient solution into the embryonic amniotic fluid [in ovo (IO) feeding] may enhance development. This study examined the effect of IO feeding on d 17.5 of incubation of carbohydrates (CHO) and beta-hydroxy-beta-methylbutyrate (HMB) on small intestinal development of chickens during the pre and posthatch periods. Results shows that 48 h post-IO feeding procedure all IO feeding treatments exhibited increased villus width and surface area compared with the control group. At d 3 posthatch the surface area of an average villi was increased by 45% for the HMB IO group and by 33% for the CHO and CHO+HMB IO groups compared with controls (noninjected fertile eggs). The activity of jejunal sucrase-isomaltase (SI) was higher (P < 0.05) 48 h after IO feeding in all the IO fed embryos, whereas at day of hatch and at d 3 the CHO+HMB IO group had the highest maltase activity (P < 0.05), which was approximately 50% greater than control embryos. These observations indicated that small intestines of IO fed hatchlings were functionally at a similar stage of development as a conventionally fed 2-d-old chick. Body weight of all IO fed hatchlings was greater than controls, and these differences (P < 0.05) were sustained until the end of the experiment (10 d). At d 10 chicks that were IO fed with CHO had BW that were 2.2% higher, whereas HMB and CHO+HMB IO fed chicks showed 5 to 6.2% BW increase, respectively, compared with controls. The current study shows that the administration of exogenous nutrients into the amnion enhanced intestinal development by increasing the size of the villi and by increasing the intestinal capacity to digest disaccharides. This advantage probably leads to higher BW in IO fed chicks.

More than two billion people are micronutrient deficient. Polished grains of popular rice varieties have concentration of approximately 2 μg g−1 iron (Fe) and 16 μg g−1 zinc (Zn). The HarvestPlus breeding programs for biofortified rice target 13 μg g−1 Fe and 28 μg g−1 Zn to reach approximately 30% of the estimated average requirement (EAR). Reports on engineering Fe content in rice have shown an increase up to 18 μg g−1 in glasshouse settings; in contrast, under field conditions, 4 μg g−1 was the highest reported concentration. Here, we report on selected transgenic events, field evaluated in two countries, showing 15 μg g−1 Fe and 45.7 μg g−1 Zn in polished grain. Rigorous selection was applied to 1,689 IR64 transgenic events for insert cleanliness and, trait and agronomic performances. Event NASFer-274 containing rice nicotianamine synthase (OsNAS2) and soybean ferritin (SferH-1) genes showed a single locus insertion without a yield penalty or altered grain quality. Endosperm Fe and Zn enrichment was visualized by X-ray fluorescence imaging. The Caco-2 cell assay indicated that Fe is bioavailable. No harmful heavy metals were detected in the grain. The trait remained stable in different genotype backgrounds.

Zinc (Zn) deficiency is a prevalent micronutrient insufficiency. Although the gut is a vital organ for Zn utilization, and Zn deficiency is associated with impaired intestinal permeability and a global decrease in gastrointestinal health, alterations in the gut microbial ecology of the host under conditions of Zn deficiency have yet to be studied. Using the broiler chicken (Gallus gallus) model, the aim of this study was to characterize distinct cecal microbiota shifts induced by chronic dietary Zn depletion. We demonstrate that Zn deficiency induces significant taxonomic alterations and decreases overall species richness and diversity, establishing a microbial profile resembling that of various other pathological states. Through metagenomic analysis, we show that predicted Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways responsible for macro- and micronutrient uptake are significantly depleted under Zn deficiency; along with concomitant decreases in beneficial short chain fatty acids, such depletions may further preclude optimal host Zn availability. We also identify several candidate microbes that may play a significant role in modulating the bioavailability and utilization of dietary Zn during prolonged deficiency. Our results are the first to characterize a unique and dysbiotic cecal microbiota during Zn deficiency, and provide evidence for such microbial perturbations as potential effectors of the Zn deficient phenotype.

BackgroundOur objective was to determine if a biofortified variety of black bean can provide more bioavailable-iron (Fe) than a standard variety. Two lines of black beans (Phaseolus-vulgaris L.), a standard (DOR500; 59μg Fe/g) and biofortified (MIB465; 88μg Fe/g) were used. The DOR500 is a common commercial variety, and the MIB465 is a line developed for higher-Fe content. Given the high prevalence of Fe-deficiency anemia worldwide, it is important to determine if Fe-biofortified black beans can provide more absorbable-Fe.MethodsBlack bean based diets were formulated to meet the nutrient requirements for the broiler (Gallus-gallus) except for Fe (dietary Fe-concentrations were 39.4±0.2 and 52.9±0.9 mg/kg diet, standard vs. biofortified, respectively). Birds (n=14) were fed the diets for 6-weeks. Hemoglobin-(Hb), liver-ferritin and Fe-related transporter/enzyme gene-expression were measured. Hemoglobin-maintenance-efficiency and total-body-Hb-Fe values were used to estimate Fe-bioavailability.ResultsHemoglobin-maintenance-efficiency values were higher (P<0.05) in the group consuming the standard-Fe beans on days 14, 21 and 28; indicating a compensatory response to lower dietary-Fe. Final total-Hb-Fe body content was higher in the biofortified vs. the standard group (26.6±0.9 and 24.4±0.8 mg, respectively; P<0.05). There were no differences in liver-ferritin or in expression of DMT-1, Dcyt-B, and ferroportin. In-vitro Fe-bioavailability assessment indicated very low Fe-bioavailability from both diets and between the two bean varieties (P>0.05). Such extremely-low in-vitro Fe-bioavailability measurement is indicative of the presence of high levels of polyphenolic-compounds that may inhibit Fe-absorption. High levels of these compounds would be expected in the black bean seed-coats.ConclusionsThe parameters of Fe-status measured in this study indicate that only a minor increase in absorbable-Fe was achieved with the higher-Fe beans. The results also raise the possibility that breeding for increased Fe-concentration elevated the levels of polyphenolic-compounds that can reduce bean Fe-bioavailability, although the higher levels of polyphenolics in the higher-Fe beans may simply be coincidental or an environmental effect. Regardless, Fe-biofortified beans remain a promising vehicle for increasing intakes of bioavailable-Fe in human populations that consume high levels of these beans as a dietary staple, and the bean polyphenol profile must be further evaluated and modified if possible in order to improve the nutritional quality of higher-Fe beans.

In nutritional studies, polyphenolic compounds are considered to be inhibitors of Fe bioavailability. Because they are presumed to act in a similar manner, total polyphenols are commonly measured via the Folin-Ciocalteu colorimetric assay. This study measured the content of polyphenolic compounds in white and black beans and examined the effect of individual polyphenols on iron uptake by Caco-2 cells. Analysis of seed coat extracts by LC-MS revealed the presence of a range of polyphenols in black bean, but no detectable polyphenols in white bean. Extracts from black bean seed coats strongly inhibited iron uptake. Examination of the eight most abundant black bean seed coat, non-anthocyanin polyphenols via Caco-2 cell assays showed that four (catechin, 3,4-dihydroxybenzoic acid, kaempferol, and kaempferol 3-glucoside) clearly promoted iron uptake and four (myricetin, myricetin 3-glucoside, quercetin, and quercetin 3-glucoside) inhibited iron uptake. The four inhibitors were present in 3-fold higher total concentration than the promoters (143 ± 7.2 vs 43.6 ± 4.4 μM), consistent with the net inhibitory effect observed for black bean seed coats. The ability of some polyphenols to promote iron uptake and the identification of specific polyphenols that inhibit Fe uptake suggest a potential for breeding bean lines with improved iron nutritional qualities.

BackgroundOur objective was to compare the capacities of biofortified and standard colored beans to deliver iron (Fe) for hemoglobin synthesis. Two isolines of large-seeded, red mottled Andean beans (Phaseolus vulgaris L.), one standard ("Low Fe") and the other biofortified ("High Fe") in Fe (49 and 71 μg Fe/g, respectively) were used. This commercial class of red mottled beans is the preferred varietal type for most of the Caribbean and Eastern and Southern Africa where almost three quarters of a million hectares are grown. Therefore it is important to know the affect of biofortification of these beans on diets that simulate human feeding studies.MethodsMaize-based diets containing the beans were formulated to meet the nutrient requirements for broiler except for Fe (Fe concentrations in the 2 diets were 42.9 ± 1.2 and 54.6 ± 0.9 mg/kg). One day old chicks (Gallus gallus) were allocated to the experimental diets (n = 12). For 4 wk, hemoglobin, feed-consumption and body-weights were measured.ResultsHemoglobin maintenance efficiencies (HME) (means ± SEM) were different between groups on days 14 and 21 of the experiment (P < 0.05). Final total body hemoglobin Fe contents were different between the standard (12.58 ± 1.0 mg {0.228 ± 0.01 μmol}) and high Fe (15.04 ± 0.65 mg {0.273 ± 0.01 μmol}) bean groups (P < 0.05). At the end of the experiment, tissue samples were collected from the intestinal duodenum and liver for further analyses. Divalent-metal-transporter-1, duodenal-cytochrome-B, and ferroportin expressions were higher and liver ferritin was lower (P < 0.05) in the standard group vs. the biofortified group. In-vitro analysis showed lower iron bioavailability in cells exposed to standard ("Low Fe") bean based diet.ConclusionsWe conclude that the in-vivo results support the in-vitro observations; biofortified colored beans contain more bioavailable-iron than standard colored beans. In addition, biofortified beans seems to be a promising vehicle for increasing intakes of bioavailable Fe in human populations that consume these beans as a dietary staple. This justifies further work on the large-seeded Andean beans which are the staple of a large-region of Africa where iron-deficiency anemia is a primary cause of infant death and poor health status.