Progress 10/01/14 to 09/30/18
Outputs
Target Audience:This project targets maize breeders and people involved in chicken husbandry. Our current work involves breeding maize that has elevated amounts of beta-cryptoxanthin and feeding it to laying hens to determine its effect on egg yolk vitamin A value. Changes/Problems:We finished our main studies with beta-cryptoxanthin and then extended the models to determine the vitamin A value of dried carrot leaves in both laying hens and Mongolian gerbils. What opportunities for training and professional development has the project provided?These studies supported at least three graduated students and many undergraduate workers during the four-year period. How have the results been disseminated to communities of interest?We have written up the results and submitted them to peer-reviewed journals. What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
We accomplished these goals and then continued our studies in laying hens and Mongolian Gerbils by performing the following studies with carrot leaves (a rich source of carotenoids): Study 1 in laying hens: The addition of food manufacturing byproducts, including carrot leaves, to animal feed is a promoted method of improving animal nutrition. The ability of dehydrated carrot leaves to improve egg yolk color and lutein and zeaxanthin (L+Z) concentration was evaluated by feeding laying hens (n = 40) white maize-based feeds fortified with two different dehydrated carrot leaves, marigold as a positive control, or no fortificant as a negative control for 28 days. After a 7-d washout period, the hens were separated into 4 groups, and eggs were collected every other day. Yolks were analyzed by using a portable colorimeter to define the color space and by Ultra-performance Liquid Chromatography to determine the carotenoid profile. Carotenoid concentration rapidly declined from d 0 to 8, confirming adequate washout conditions. The white maize negative control (WM) d 28 lutein concentration (3.59 +/- 0.51 nmol/g) was significantly less than orange-carrot leaf-treated (OCL) (5.34 +/- 0.36 nmol/g) and red-carrot leaf-treated hens (RCL) (5.92 +/- 1.00 nmol/g) in addition to the marigold-treated hens (MG). However, MG was significantly higher than both leaf-treated groups. From d 8 (3.93 +/- 0.74 nmol/g) to d 28 (9.32 +/- 1.66 nmol/g), MG had the largest increase in lutein and was the only treatment to surpass d 0 initial concentrations (8.50 1.64 nmol/g). A similar trend was observed for zeaxanthin and was reflected in the color space. Study 2 in Mongolian Gerbils: After 3-wk depletion, baseline Mongolian gerbils were killed (n = 6), and remaining gerbils (n = 60) were divided into 6 groups to receive four carrot leaf-fortified feeds equalized for beta-carotene equivalents (bCE), or VA-deficient feed with (VA+) or without (VA-) retinyl acetate supplements. Carrot-leaf powder from four different carrot plants with differing alpha:beta-carotene were used. After 4-wk, gerbils were killed. Serum and tissues were collected and analyzed for retinoids by HPLC. One-way ANOVA was used with multiple group comparisons for variables of interest. VA+ had higher total liver VA (0.91 ± 0.29 mmol) than all other groups (P < 0.03). The carrot leaf-treatments did not differ from baseline (0.55 ± 0.09 mmol). VA- (0.40 ± 0.23 mmol) did not differ from leaf-fed groups, but 30% became VA deficient (defined as <0.1 micromol VA/g liver). alpha-Retinol accumulated in livers and lungs and was correlated to total alpha-carotene consumption (R2 = 0.83 and 0.88, respectively; P < 0.0001). Bioefficacy factors ranged from 4.2 to 6.2 mg bCE to 1 mg retinol. Carrot leaves maintained VA status and prevented deficiency in gerbils regardless of alpha:beta-carotene. The bioconversion of provitamin A carotenoids from carrot leaves to retinol was similar to that of other green leafy vegetables, making the addition of carrot leaves to human food and animal feeds a viable method to improve dietary carotenoid content.
Publications
- Type:
Journal Articles
Status:
Submitted
Year Published:
2019
Citation:
T. J. Titcomb,* M. S. Kaeppler,* M. E. Cook,*,P. W. Simon* and S. A. Tanumihardjo. Carrot leaves improve color and xanthophyll content of egg yolk in laying hens but are not as effective as commercially available marigold fortificant. Poultry Science (submitted).
- Type:
Journal Articles
Status:
Under Review
Year Published:
2019
Citation:
Tyler J. Titcomb, Mikayla S. Kaeppler, Sof�a Beatriz Sandoval Cates, Jamie M. Shannon, Philipp W. Simon, Sherry A. Tanumihardjo. Carrot Leaves Maintain Liver Vitamin A Concentrations in Male Mongolian Gerbils Regardless of the alpha- to beta-Carotene Ratio when beta-Carotene Equivalents are Equalized. J. Nutrition (In review).
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Progress 10/01/16 to 09/30/17
Outputs
Target Audience:This project targets maize breeders and people involved in chicken husbandry. Our current work involves breeding maize that has elevated amounts of beta-cryptoxanthin and feeding it to laying hens to determine its effect on egg yolk vitamin A value. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest?We continue to work closely with maize breeders at the Center for Maize and Wheat Improvement in Texcoco, Mexico. The graduate student finishing the studies is looking at different levels of vitamin A on provitamin A bioconversion, successfully completed her MS degree. What do you plan to do during the next reporting period to accomplish the goals?We will be resubmitting the manuscript on Specific Aim 3 in February 2018. We will be finishing the manuscript for Specific Aim 2 by the end of the grant period.
Impacts
What was accomplished under these goals?
We finished the work on Specific Aim 3 Cryptoxanthin-biofortified eggs were produced from chickens who we fed biofortified orange or tangerine-fortified maize feeds and fed to Mongolian gerbils. Gerbils (n = 57) were fed VA-deficient feed 28 d. After baseline (n = 7), treatments (n = 10/group) included oil control (VA-); 16.7% orange maize-biofortified, tangerine-fortified, or white-maize egg feeds; or retinyl acetate in oil (VA+) matched to daily preformed retinol intake from the eggs for 30 d. Liver fat (P < 0.0001) and cholesterol (P < 0.004) were higher in the egg-fed groups. Liver fatty acid profiles reflected feed, but not retinyl esters. Liver retinol concentration (0.13 + 0.03 µmol/g) and total reserves (0.52 + 0.12 µmol) were highest in gerbils fed orange-biofortified eggs than all other groups (P < 0.05). Liver retinol was similar among VA+, tangerine-egg, and white-egg-fed gerbils but total retinol reserves were higher in tangerine-egg fed than VA+, VA-, and baseline (P < 0.05). The VA- group was deficient (0.018 + 0.010 µmol retinol/g liver). Eggs enhanced VA status but b-cryptoxanthin biofortified eggs prevented deficiency. Biofortified maize can enhance VA status when consumed directly or through products from livestock fed biofortified maize. After this study we performed two other gerbil studies to look at the influence of preformed vitamin A on beta-carotene bioconversion. This was done to evaluate Specific Aim 2's objective. Two studies in Mongolian gerbils simulated the condition of exposure to multiple VA interventions. The effects of provitamin A consumption from biofortified orange maize and carrots, and VA consumption from preformed VA fortificant on VA status were investigated. Study 1 was a 2x2x2 factorial (n = 85) with biofortified high-beta-carotene maize, orange carrots, and VA fortification at 50% of estimated gerbil needs, compared with respective white and VA-free controls. Study 2 was a 2x3 factorial (n = 66) evaluating the effects of orange carrot and excessive VA consumption through fortification at 100 and 200% of estimated needs. Both studies utilized a 2-wk VA-depletion phase, baseline evaluation, 9-wk treatment phase, and determined VA status with liver stores. In Study 1, liver retinol concentrations (in micromol/g liver) were significantly higher than baseline (0.232 ± 0.069) and respective controls in the orange carrot group (0.686 ± 0.123) and orange maize group (0.523 ± 0.208). VA+ fortificant alone did not significantly improve VA status (0.107 ± 0.053). In Study 2, orange carrot addition to feeds significantly enhanced liver stores (0.854 ± 0.235) relative to baseline (0.432 ± 0.136), but VA fortificant alone (0.415 ± 0.212) did not. Serum retinol was not affected by dietary VA or liver reserves in either study. Orange carrots and biofortified maize prevented VA deficiency as single interventions. On the background of adequate dietary intake of provitamin A carotenoids from carrots, VA fortification caused excessive and hypervitaminotic liver stores in this animal model.
Publications
- Type:
Journal Articles
Status:
Accepted
Year Published:
2017
Citation:
Sowa M, Yu J, Palacios-Rojas N, Goltz SR, Howe JA, Davis CR, Rocheford T, Tanumihardjo SA. Retention of carotenoids in biofortified maize flour and beta-cryptoxanthin-enhanced eggs after household cooking. ACS Omega. 2017;doi.org/10.1021/acsomega.7b01202.
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Progress 10/01/15 to 09/30/16
Outputs
Target Audience:The target audience for this work is consumers that are interesting in improving their health. We are working on biofortified foods. We are currently showing proof of concept in both chickens and gerbils. Changes/Problems:We were originally planning to do another chicken feeding study in 2017; however, the specialty maize that was being grown in Mexico for this purpose was infected with a virus. We may try to isolate beta-cryptoxanthin from tangerine peel and compare it with the peel itself. We are currently collecting peels. It may be difficult to get enough semi-purifed beta-cryptoxanthin to do a chicken study. However, we are planning a gerbil study this year also. What opportunities for training and professional development has the project provided?The project has provided support for graduate students, one of which recently finished his PhD and is currently post-docing at Cornell University. We have recently hired two new students who are working with the biofortified maize and will be working with chickens and gerbils in the near future. How have the results been disseminated to communities of interest?We have given several presentations over the years to a variety of audiences. What do you plan to do during the next reporting period to accomplish the goals?We are getting very close at finishing the third AIM paper and will be submitting it in 2017. We will also do one more chicken study and gerbil study before the closing of this grant.
Impacts
What was accomplished under these goals?
We have finished two studies in regard to AIM 2 in Mongolian gerbils. Two Mongolian gerbil studies (male, 29-35 d) investigated the interaction of maize milling methods (i.e., whole-grain vs. refined) with conventionally bred, provitamin A-biofortified (orange) or white maize on vitamin A and zinc statuses. It was important to show these milling effects because different population groups have various preparation methods. This study was a spin off of our other work in this area and was recently accepted for publication. Milling did not affect carotenoid bioefficacy but it did affect zinc status in these animals. Another related study was published in 2016. This was a substudy of a study from a visiting post-doc. The graduate student did a secondary analysis looking at the changes in natural enrichment of 13C with biofortified maize feeding to Mongolian gerbils. This showed innovativeness on the student's part to realize he could get something publishable on samples that had been collected for a different purpose. From this study, we were able to show shifts in the natural abundance of 13C in the retinol circulating and stores in the animals. We have also finished the work for AIM 3 by feeding biofortified maize to chickens (AIM 1 published) and then feeding the eggs to gerbils. This manuscript is almost in its final form.
Publications
- Type:
Journal Articles
Status:
Accepted
Year Published:
2017
Citation:
Gannon BM, Pixley KV, Tanumihardjo SA. Maize Milling Method affects Growth and Zinc Status, but not Provitamin A Carotenoid Bioefficacy, in Male Mongolian Gerbils. Journal of Nutrition 2017:(in press).
- Type:
Journal Articles
Status:
Published
Year Published:
2016
Citation:
Gannon BM, Pungarcher I, Mourao L, Davis CR, Simon P, Pixley KV, Tanumihardjo SA. 13C natural abundance of serum retinol is a novel biomarker for evaluating provitamin A carotenoid-biofortified maize consumption in male Mongolian gerbils. J Nutr. 2016;146(7):1290-7.
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Progress 10/01/14 to 09/30/15
Outputs
Target Audience:This is a multidisciplinary project. We are targeting those that are interested in poultry feeds and enhancement of human health by producing biofortified eggs. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?This study supported a graduate student and many undergraduate student researchers. We will continue these studies with enhancing the beta-cryptoxanthin concentrations in the egg yolks. We have described the comparison with tangerine peel powder. How have the results been disseminated to communities of interest?We have published one paper and are currently working on another one. The results of feeding eggs to gerbils will likely have more outcomes than improving vitamin A status. They may also be a unique model for cholesterol feeding studies. What do you plan to do during the next reporting period to accomplish the goals?We are hoping to finish the Mongolian gerbil paper. We also did another feeding study with biofortified maize directly to gerbils and have some spin off studies related to that.
Impacts
What was accomplished under these goals?
Chickens have the ability to deposit xanthophyll carotenoids into egg yolk, which gives eggs their bright yellow hue. The xanthophyll β-cryptoxanthin provides vitamin A and has other purported health benefits.In line with objective 1, high β-cryptoxanthin chicken feeds were prepared to investigate egg yolk color changes, yolk carotenoid enhancement, and resultant hen vitamin A status.After a 10-d washout period on white maize feed, chickens (n = 8/group) were fed high β-cryptoxanthin biofortified (orange) maize, tangerine peel-fortified white maize, lutein-fortified yellow maize (industry standard), or white maize only (negative control) for 40 d. Color, retinol, and carotenoid analyses were performed on egg yolks periodically during the study. Hens were killed on d 50 and liver, serum, and skin were analyzed for retinol equivalents and carotenoids. Egg yolks from chickens fed the orange maize feed had significantly different color scores for the L- (light/dark), a- (red/green), and b- (yellow/blue) scales, and had significantly higher β-cryptoxanthin concentrations (8.55 + 1.45 nmol/g from d 24-50) than other treatment groups (P < 0.0001 for all comparisons). Retinol egg yolk concentrations were similar among treatment groups and decreased with time (P < 0.0001). Hens fed the orange maize had significantly higher liver retinol concentrations (0.53 + 0.20 µmol/g) and reserves (18.9 + 7.12 µmol) than other groups (P < 0.0001). Serum retinol concentration was highest in chickens fed the tangerine fortificant (1.94 + 0.14 mmol/L, P < 0.0001). No differences were observed in chicken skin color assessment among groups, but β-cryptoxanthin concentration was highest in the orange maize-fed group (P = 0.002).In conclusion,consumption of β-cryptoxanthin from biofortified maize by chickens enhances egg yolk color and β-cryptoxanthin concentrations. β-Cryptoxanthin-biofortified eggs could be another choice for consumers of lutein-fortified eggs, providing both vitamin A and carotenoids further supporting eggs' status as a functional food. In line with objective 3, we have subsequently fed the biofortified egg yolk to Mongolian gerbils. That publication is currently being finalized.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2014
Citation:
Heying E, Tanumihardjo JP, Verdan V, Cook ME, Palacios-Rojas N, Tanumihardjo SA. Biofortified orange maize enhances beta-cryptoxanthin concentrations in egg yolks of laying hens better than tangerine peel fortificant. J Agric Food Chem. 2014;62(49):11892-900.
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